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1.
J Transl Med ; 22(1): 493, 2024 May 24.
Artículo en Inglés | MEDLINE | ID: mdl-38789992

RESUMEN

BACKGROUND: Autologous bone grafting is the standard treatment for the surgical management of atrophic nonunion of long bones. Other solutions, such as bone marrow mesenchymal stem cells (BM-MSC) combined with phospho-calcium material, have also been used. Here we evaluate the safety and early efficacy of a novel procedure using autologous or allogenic adipose tissue mesenchymal stromal cells (AT-MSC) seeded in a patented tricalcium phosphate-based biomaterial for the treatment of bone regeneration in cases of atrophic nonunion. METHODS: This was a prospective, multicentric, open-label, phase 2 clinical trial of patients with atrophic nonunion of long bones. Biografts of autologous or allogenic AT-MSC combined with a phosphate substrate were manufactured prior to the surgical procedures. The primary efficacy was measured 6 months after surgery, but patients were followed for 12 months after surgery and a further year out of the scope of the study. All adverse events were recorded. This cohort was compared with a historical cohort of 14 cases treated by the same research team with autologous BM-MSC. RESULTS: A total of 12 patients with atrophic nonunion of long bones were included. The mean (SD) age was 41.2 (12.1) years and 66.7% were men. Bone healing was achieved in 10 of the 12 cases (83%) treated with the AT-MSC biografts, a percentage of healing similar (11 of the 14 cases, 79%) to that achieved in patients treated with autologous BM-MSC. Overall, two adverse events, in the same patient, were considered related to the procedure. CONCLUSIONS: The results of this study suggest that AT-MSC biografts are safe for the treatment of bone regeneration in cases of atrophic nonunion and reach high healing rates. TRIAL REGISTRATION: Study registered with EUDRA-CT (2013-000930-37) and ClinicalTrials.gov (NCT02483364).


Asunto(s)
Tejido Adiposo , Materiales Biocompatibles , Fosfatos de Calcio , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , Trasplante Autólogo , Humanos , Fosfatos de Calcio/farmacología , Fosfatos de Calcio/uso terapéutico , Células Madre Mesenquimatosas/citología , Masculino , Femenino , Persona de Mediana Edad , Tejido Adiposo/citología , Adulto , Trasplante Homólogo , Resultado del Tratamiento , Atrofia , Estudios Prospectivos
2.
Mol Biol Rep ; 51(1): 838, 2024 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-39042226

RESUMEN

BACKGROUND: Bioglass materials have gained significant attention in the field of tissue engineering due to their osteoinductive and biocompatible properties that promote bone cell differentiation. In this study, a novel composite scaffold was developed using a sol-gel technique to combine bioglass (BG) 58 S with a poly L-lactic acid (PLLA). METHODS AND RESULTS: The physiochemical properties, morphology, and osteoinductive potential of the scaffolds were investigated by X-ray diffraction analysis, scanning electron microscopy, and Fourier-transform infrared spectroscopy. The results showed that the SiO2-CaO-P2O5 system was successfully synthesized by the sol-gel method. The PLLA scaffolds containing BG was found to be osteoinductive and promoted mineralization, as demonstrated by calcium deposition assay, upregulation of alkaline phosphatase enzyme activity, and Alizarin red staining data. CONCLUSIONS: These in vitro studies suggest that composite scaffolds incorporating hBMSCs are a promising substitute material to be implemented in bone tissue engineering. The PLLA/BG scaffolds promote osteogenesis and support the differentiation of bone cells, such as osteoblasts, due to their osteoinductive properties.


Asunto(s)
Materiales Biocompatibles , Diferenciación Celular , Cerámica , Osteogénesis , Poliésteres , Ingeniería de Tejidos , Andamios del Tejido , Poliésteres/química , Andamios del Tejido/química , Cerámica/química , Cerámica/farmacología , Ingeniería de Tejidos/métodos , Materiales Biocompatibles/farmacología , Materiales Biocompatibles/química , Osteogénesis/efectos de los fármacos , Humanos , Diferenciación Celular/efectos de los fármacos , Regeneración Ósea/efectos de los fármacos , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Difracción de Rayos X , Huesos/efectos de los fármacos , Huesos/metabolismo , Fosfatasa Alcalina/metabolismo , Microscopía Electrónica de Rastreo
3.
J Transl Med ; 21(1): 781, 2023 11 04.
Artículo en Inglés | MEDLINE | ID: mdl-37925419

RESUMEN

BACKGROUND: Diabetes mellitus (DM) and periodontitis are two prevalent diseases with mutual influence. Accumulation of advanced glycation end products (AGEs) in hyperglycemia may impair cell function and worsen periodontal conditions. N6-methyladenosine (m6A) is an important post-transcriptional modification in RNAs that regulates cell fate determinant and progression of diseases. However, whether m6A methylation participates in the process of periodontitis with diabetes is unclear. Thus, we aimed to investigate the effects of AGEs on bone marrow mesenchymal stem cells (BMSCs), elucidate the m6A modification mechanism in diabetes-associated periodontitis. METHODS: Periodontitis with diabetes were established by high-fat diet/streptozotocin injection and silk ligation. M6A modifications in alveolar bone were demonstrated by RNA immunoprecipitation sequence. BMSCs treated with AGEs, fat mass and obesity associated (FTO) protein knockdown and sclerostin (SOST) interference were evaluated by quantitative polymerase chain reaction, western blot, immunofluorescence, alkaline phosphatase and Alizarin red S staining. RESULTS: Diabetes damaged alveolar bone regeneration was validated in vivo. In vitro experiments showed AGEs inhibited BMSCs osteogenesis and influenced the FTO expression and m6A level in total RNA. FTO knockdown increased the m6A levels and reversed the AGE-induced inhibition of BMSCs differentiation. Mechanically, FTO regulated m6A modification on SOST transcripts, and AGEs affected the binding of FTO to SOST transcripts. FTO knockdown accelerated the degradation of SOST mRNA in presence of AGEs. Interference with SOST expression in AGE-treated BMSCs partially rescued the osteogenesis by activating Wnt Signaling. CONCLUSIONS: AGEs impaired BMSCs osteogenesis by regulating SOST in an m6A-dependent manner, presenting a promising method for bone regeneration treatment of periodontitis with diabetes.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato , Diabetes Mellitus , Células Madre Mesenquimatosas , Periodontitis , Humanos , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato/genética , Células de la Médula Ósea/metabolismo , Diferenciación Celular , Células Cultivadas , Productos Finales de Glicación Avanzada/farmacología , Osteogénesis , Periodontitis/genética , ARN/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética
4.
Cell Tissue Bank ; 24(3): 663-681, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-36622494

RESUMEN

Skeletal problems are an increasing issue due to the increase in the global aging population. Different statistics reports show that today, the global population is aging that results in skeletal problems, increased health system costs, and even higher mortality associated with skeletal problems. Common treatments such as surgery and bone grafts are not always effective and in some cases, they can even cause secondary problems such as infections or improper repair. Cell therapy is a method that can be utilized along with common treatments independently. Mesenchymal stem cells (MSCs) are a very important and efficient source in terms of different diseases, especially bone problems. These cells are present in different tissues such as bone marrow, adipose tissue, umbilical cord, placenta, dental pulp, peripheral blood, amniotic fluid and others. Among the types of MSCs, bone marrow mesenchymal stem cells (BMMSCs) are the most widely used source of these cells, which have appeared to be very effective and promising in terms of skeletal diseases, especially compared to the other sources of MSCs. This study focuses on the specific potential and content of BMMSCs from which the specific capacity of these cells originates, and compares their osteogenic potential with other types of MSCs, and also the future directions in the application of BMMSCs as a source for cell therapy.


Asunto(s)
Células Madre Mesenquimatosas , Osteogénesis , Embarazo , Femenino , Humanos , Huesos , Placenta , Células Cultivadas , Células Madre Mesenquimatosas/metabolismo , Células de la Médula Ósea/metabolismo , Diferenciación Celular
5.
BMC Oral Health ; 23(1): 247, 2023 04 28.
Artículo en Inglés | MEDLINE | ID: mdl-37118728

RESUMEN

OBJECTIVES: Dentin, the bulk material of the tooth, resemble the bone's chemical composition and is considered a valuable bone substitute. In the current study, we assessed the cytotoxicity and osteogenic potential of demineralized dentin matrix (DDM) in comparison to HA nanoparticles (n-HA) on bone marrow mesenchymal stem cells (BMMSCs) using a hydrogel formulation. MATERIALS AND METHODS: Human extracted teeth were minced into particles and treated via chemical demineralization using ethylene diamine tetra-acetic acid solution (EDTA) to produce DDM particles. DDM and n-HA particles were added to the sodium alginate then, the combination was dripped into a 5% (w/v) calcium chloride solution to obtain DDM hydrogel (DDMH) or nano-hydroxyapatite hydrogel (NHH). The particles were evaluated by dynamic light scattering (DLS) and the hydrogels were evaluated via scanning electron microscope (SEM). BMMSCs were treated with different hydrogel concentrations (25%, 50%, 75% and neat/100%) and cell viability was evaluated using MTT assay after 72 h of culture. Collagen-I (COL-I) gene expression was studied with real-time quantitative polymerase chain reaction (RT-qPCR) after 3 weeks of culture and alkaline phosphatase (ALP) activity was assessed using enzyme-linked immune sorbent assay (ELISA) over 7th, 10th, 14th and 21st days of culture. BMMSCs seeded in a complete culture medium were used as controls. One-way ANOVA was utilized to measure the significant differences in the tested groups. RESULTS: DLS measurements revealed that DDM and n-HA particles had negative values of zeta potential. SEM micrographs showed a porous microstructure of the tested hydrogels. The viability results revealed that 100% concentrations of either DDMH or NHH were cytotoxic to BMMSCs after 72 h of culture. However, the cytotoxicity of 25% and 50% concentrations of DDMH were not statistically significant compared to the control group. RT-qPCR showed that COL-I gene expression was significantly upregulated in BMMSCs cultured with 50% DDMH compared to all other treated or control groups (P < 0.01). ELISA analysis revealed that ALP level was significantly increased in the groups treated with 50% DDMH compared to 50% NHH after 21 days in culture (P < 0.001). CONCLUSION: The injectable hydrogel containing demineralized dentin matrix was successfully formulated. DDMH has a porous structure and has been shown to provide a supporting matrix for the viability and differentiation of BMMSCs. A 50% concentration of DDMH was revealed to be not cytotoxic to BMMSCs and may have a great potential to promote bone formation ability.


Asunto(s)
Hidrogeles , Células Madre Mesenquimatosas , Humanos , Hidrogeles/farmacología , Hidrogeles/análisis , Hidrogeles/química , Osteogénesis , Dentina/química , Durapatita/farmacología , Durapatita/química , Colágeno Tipo I , Diferenciación Celular
6.
BMC Musculoskelet Disord ; 23(1): 557, 2022 Jun 09.
Artículo en Inglés | MEDLINE | ID: mdl-35681160

RESUMEN

BACKGROUND: The choice of bone substitutes for the treatment of infected bone defects (IBDs) has attracted the attention of surgeons for years. However, single-stage bioabsorbable materials that are used as carriers for antibiotic release, as well as scaffolds for BMSC sheets, need further exploration. Our study was designed to investigate the effect of vancomycin-loaded calcium sulfate hemihydrate/nanohydroxyapatite/carboxymethyl chitosan (CSH/n-HA/CMCS) hydrogels combined with BMSC sheets as bone substitutes for the treatment of IBDs. METHODS: BMSCs were harvested and cultured into cell sheets. After the successful establishment of an animal model with chronic osteomyelitis, 48 New Zealand white rabbits were randomly divided into 4 groups. Animals in Group A were treated with thorough debridement as a control. Group B was treated with BMSC sheets. CSH/n-HA/CMCS hydrogels were implanted in the treatment of Group C, and Group D was treated with CSH/n-HA/CMCS+BMSC sheets. Gross observation and micro-CT 3D reconstruction were performed to assess the osteogenic and infection elimination abilities of the treatment materials. Histological staining (haematoxylin and eosin and Van Gieson) was used to observe inflammatory cell infiltration and the formation of collagen fibres at 4, 8, and 12 weeks after implantation. RESULTS: The bone defects of the control group were not repaired at 12 weeks, as chronic osteomyelitis was still observed. HE staining showed a large amount of inflammatory cell infiltration around the tissue, and VG staining showed no new collagen fibres formation. In the BMSC sheet group, although new bone formation was observed by gross observation and micro-CT scanning, infection was not effectively controlled due to unfilled cavities. Some neutrophils and only a small amount of collagen fibres could be observed. Both the hydrogel and hydrogel/BMSCs groups achieved satisfactory repair effects and infection control. Micro-CT 3D reconstruction at 4 weeks showed that the hydrogel/BMSC sheet group had higher reconstruction efficiency and better bone modelling with normal morphology. HE staining showed little aggregation of inflammatory cells, and VG staining showed a large number of new collagen fibres. CONCLUSIONS: Our preliminary results suggested that compared to a single material, the novel antibiotic-impregnated hydrogels acted as superior scaffolds for BMSC sheets and excellent antibiotic vectors against infection, which provided a basis for applying tissue engineering technology to the treatment of chronic osteomyelitis.


Asunto(s)
Sustitutos de Huesos , Quitosano , Osteomielitis , Animales , Conejos , Antibacterianos , Sulfato de Calcio , Colágeno , Hidrogeles , Osteogénesis , Osteomielitis/tratamiento farmacológico , Andamios del Tejido , Vancomicina
7.
Oral Dis ; 27(6): 1551-1563, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-33078488

RESUMEN

OBJECTIVE: To determine whether risk factors other than hyperglycemia lead to failed osseointegration in patients with type 2 diabetes mellitus (T2DM) during the healing period. MATERIALS AND METHODS: We compared the success rates between patients with and without T2DM during the healing period at our center. Bone marrow mesenchymal stem cells (BMSCs) were cultured from subjects. Proteomics was used to detect differentially expressed proteins (DEPs) among the DM failure (DM-F), DM success (DM-S), and control (Con) groups. The correlation between the expression levels of nine target DEPs and medium glucose concentrations was investigated. RESULTS: Higher failure rates were observed in the T2DM patients. Fifty-two DEPs were found between the DM-F and DM-S groups. Seventy-three DEPs were found between the DM-F and Con groups. Forty-three DEPs were found between the DM-S and Con groups. Five target DEPs were expressed at the same levels in the medium with different glucose concentrations. Gene ontology annotation and functional enrichment analysis suggest that the DEPs detected in the DM-F group may affect the biological function and regulatory potential of BMSCs. CONCLUSIONS: The DEPs detected in the DM-F group can be intervention targets for to prevent implant failure in T2DM patients. Risk factors besides hyperglycemia may affect osseointegration during healing period.


Asunto(s)
Implantes Dentales , Diabetes Mellitus Tipo 2 , Hiperglucemia , Diabetes Mellitus Tipo 2/complicaciones , Humanos , Hiperglucemia/complicaciones , Oseointegración , Factores de Riesgo
8.
Biochem Biophys Res Commun ; 531(3): 290-296, 2020 10 20.
Artículo en Inglés | MEDLINE | ID: mdl-32800542

RESUMEN

Dental implant is the most effective way to repair the defect or absence of dentition. Currently, the modification in titanium surface properties has become a hot topic in the study of oral implantology. However, more suitable titanium surface coating still needs to be further explored. We prepared the nerve growth factor-chondroitin sulfate/hydroxyapatite (NGF-CS/HA)-coating composite titanium by modified biomimetic method. We also observed the surface morphology, thickness, surface adhesion and component analysis of NGF-CS/HA-coating composite titanium by scanning electron microscope, and the release of NGF was also identified via ELISA assay. Besides, the identification of bone marrow mesenchymal stem cells (BMSCs) was conducted through alizarin red staining, oil red O staining and fluorescence detection. and the osteogenesis differentiation and neuronal differentiation-related genes were determined by RT-qPCR assay. The surface of NGF-CS/HA coating with the 65.4 ± 6.4 µm thickness presented a porous network, and the main components of NGF-CS/HA coating were Ti and HA, and maintained the activity and release of NGF. Besides, we successfully obtained and identified BMSCs, and proved that NGF-CS/HA-coating composite titanium could notably upregulated the expression levels of the osteogenesis differentiation and neuronal differentiation-related genes and proteins in BMSCs. In conclusion, NGF-CS/HA-coating composite titanium has significant promoting effects on the differentiation of BMSCs into osteoblast and neural cells.


Asunto(s)
Diferenciación Celular/efectos de los fármacos , Sulfatos de Condroitina/farmacología , Materiales Biocompatibles Revestidos/farmacología , Células Madre Mesenquimatosas/citología , Factor de Crecimiento Nervioso/farmacología , Neuronas/citología , Osteoblastos/citología , Titanio/farmacología , Fosfatasa Alcalina/metabolismo , Animales , Calcio/metabolismo , Adhesión Celular/efectos de los fármacos , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Neurogénesis/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Propiedades de Superficie
9.
Connect Tissue Res ; 61(6): 577-585, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-31305177

RESUMEN

Purpose: Human jaw bone marrow mesenchymal stem cells (h-JBMMSCs) are multipotent progenitor cells with osteogenic differentiation potential. MicroRNAs (miRNAs) have emerged as crucial modulators of osteoblast differentiation. In this study, we focus on the role of miR-145 and its target protein in osteoblast differentiation of h-JBMMSCs. Materials and Methods: h-JBMMSCs were isolated and cultured in osteogenic medium. miR-145 mimics and inhibitors were used to elevate and inhibit miR-145 expression, respectively. Osteogenic differentiation was determined by Alkaline phosphatase (ALP) and Alizarin red S (ARS) staining, and osteogenic marker detection using quantitative real-time reverse transcription PCR (qRT-PCR) assay. Bioinformatic analysis and luciferase reporter assay were used to identify the target gene of miR-145. Results: MiR-145 was down-regulated during osteogenesis of h-JBMMSCs. Inhibition of miR-145 promoted osteogenic differentiation of h-JBMMSCs, revealed by enhanced activity of alkaline phosphatase (ALP), greater mineralisation, and increased expression levels of the osteogenic markers, such as Runt-related transcription factor 2 (RUNX2), Osterix (OSX), ALP and COL1A1. MiR-145 could negatively regulate semaphorin3A (SEMA3A), which acts as a positive regulator of osteogenesis. MiR-145 inhibitor induced osteogenesis could be partially attenuated by SEMA3A siRNA treatment in h-JBMMSCs. Conclusions: Our data show that miR-145 directly targets SEMA3A, and also suggest miR-145 as a suppressor, plays an important role in the osteogenic differentiation of h-JBMMSCs.


Asunto(s)
Células de la Médula Ósea/citología , Diferenciación Celular/genética , Maxilares/citología , Células Madre Mesenquimatosas/citología , MicroARNs/metabolismo , Osteogénesis/genética , Semaforina-3A/metabolismo , Secuencia de Bases , Regulación hacia Abajo/genética , Células HEK293 , Humanos , Células Madre Mesenquimatosas/metabolismo , MicroARNs/genética
10.
J Musculoskelet Neuronal Interact ; 20(1): 142-148, 2020 03 03.
Artículo en Inglés | MEDLINE | ID: mdl-32131379

RESUMEN

OBJECTIVE: to investigate the combined construction of injectable tissue-engineered bone with calcium phosphate bone cement composite (CPC) and bone marrow mesenchymal stem cells (BMMSCs). METHODS: The proliferation activity of BMMSCs encapsulated was detected by CCK8 method on the 7th day after its self-coagulation by CPC. qRT-PCR was used to detect the expressions of mRNA. The microcapsules of BMMSCs combined with CPC were completely filled in the defect site in the experimental group, and the control group not filled. The two groups were sutured and routinely reared, double upper limb X-ray examination performed after operation. RESULTS: Those of two groups were on the rise over time, which were higher at the 1st, 3rd, 5th and 7th days than those at the previous time points (all P<0.05). The relative expressions of ALP and CALCR at the 7th day were higher than those at the day in BMMSCs combined with the CPC group and BMMSCs group (all P<0.05). The relative expression of CALCR was significantly higher in BMMSCs combined with the CPC group than that in the BMMSCs group on the 7th day (P<0.05). CONCLUSION: With good cell activity and biological activity, the combined construction of the tissue-engineered bone with BMMSCs and CPC can be used as an ideal treatment material for bone tissue repair and connection.


Asunto(s)
Cementos para Huesos/farmacología , Trasplante de Células Madre Mesenquimatosas/métodos , Radio (Anatomía)/diagnóstico por imagen , Radio (Anatomía)/lesiones , Ingeniería de Tejidos/métodos , Animales , Fosfatos de Calcio/farmacología , Células Cultivadas , Terapia Combinada/métodos , Células Madre Mesenquimatosas/fisiología , Conejos , Radio (Anatomía)/efectos de los fármacos
11.
Dent Traumatol ; 36(3): 278-284, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31797525

RESUMEN

BACKGROUND/AIMS: Lacerations of the oral mucosa and fractures of alveolar processes commonly occur in traumatic dental injuries (TDIs). Impaired wound healing and tissue regeneration have severe consequences on the quality of life. Bone marrow mesenchymal stem cells (BMMSCs) possess the ability of self-renewal and multipotential differentiation. Treatment with low-level sodium fluoride (NaF) has emerged as a promising approach to enhance wound repair. The aim of this study was to assess the effects of low-level NaF on soft tissue healing and on the proliferation, migration and extracellular matrix synthesis of BMMSCs. MATERIAL AND METHODS: BMMSCs derived from mice were treated with 50 µM, 500 µM, or 5 mM NaF for 12, 24, and 48 hours, and cell proliferation was assessed by the MTS assay. Cell motility was detected at 12 and 24 hours by a wound healing assay, and osteoblastic differentiation for 21 days by 1% Alizarin Red S staining in 50 µM NaF-treated BMMSCs. Gene expression of Runx2 and Osteocalcin was evaluated by quantitative real-time PCR. An experimental rat skin wound model was employed, and levels of c-Myc, Ki67, fibronectin, and vimentin were assessed by immunohistochemistry. RESULTS: There was a significant induction in the proliferation and migration of BMMSCs treated with 50 µM NaF. The expression of Ki67 and c-Myc protein was increased in tissues treated with 50 µM NaF, and the expression of fibronectin and vimentin in the 50 µM NaF-treated tissues was stimulated. Alizarin Red staining revealed enhanced mineralization in 50 µM NaF-treated BMMSCs with increased expression of Runx2 and Osteocalcin, indicating their upregulated osteogenic differentiation. CONCLUSION: Low-level NaF could promote soft tissue healing and hard tissue regeneration.


Asunto(s)
Células Madre Mesenquimatosas , Osteogénesis , Animales , Células de la Médula Ósea , Diferenciación Celular , Células Cultivadas , Ratones , Calidad de Vida , Ratas , Fluoruro de Sodio/farmacología , Cicatrización de Heridas
12.
Med J Armed Forces India ; 76(2): 172-179, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32476715

RESUMEN

BACKGROUND: Considering the limitations in isolating Bone Marrow Mesenchymal Stem Cells (BMSCs), alternate sources of Mesenchymal Stem Cells (MSCs) are being intensely investigated. This study evaluated dental pulp MSCs (DP-MSCs) isolated from orthodontically extracted premolar teeth from a bone tissue engineering perspective. METHODS: MSCs isolated from premolar teeth pulp were cultured and studied using BMSCs as the control. Flow cytometry analysis was performed for the positive and negative MSC markers. Multilineage differentiation focusing on bone regeneration was evaluated by specific growth induction culturing media and by alkaline phosphatase (ALP) activity. Data were compared by repeated measurement analysis of variance and Student's t-test at a p value <0.05. RESULTS: Proliferation rate, population doubling time, and colony formation of DP-MSCs were significantly higher (p < 0.001) than BMSCs. More than 85% of DP-MSCs expressed CD44, CD73, CD90, CD105, and CD166. Negative reaction was found for CD11b CD33, CD34, and CD45. Positive reaction was displayed by 7.2% of cells for early MSC marker, Stro-1. Both the cell populations differentiated into adipogenic, osteogenic, and chondrogenic lineages, with adequate ALP expression. CONCLUSION: Because DP-MSCs from orthodontic premolars hold a neural crest/ectomesenchymal ancestry, its prudent growth characteristics and multilineage differentiation open up exciting options in craniofacial tissue engineering.

13.
Adv Exp Med Biol ; 1144: 101-121, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30725365

RESUMEN

Oromaxillofacial tissues (OMT) are composed of tooth and bone, together with nerves and blood vessels. Such a composite material is a huge source for mesenchymal stem cells (MSCs) that can be obtained with ease from extracted teeth, teeth structures and socket blood, flapped gingiva tissue, and mandibular/maxillar bone marrow. They offer a biological answer for restoring damaged dental tissues such as the regeneration of alveolar bone, prevention of pulp tissue defects, and dental structures. Dental tissue-derived mesenchymal stem cells share properties with bone marrow-derived mesenchymal stem cells and there is a considerable potential for these cells to be used in different stem cell-based therapies, such as bone and nerve regeneration. Dental pulp tissue might be a very good source for neurological disorders whereas gingiva-derived mesenchymal stem cells could be a good immune modulatory/suppressive mediators. OMT-MSCs is also promising candidates for regeneration of orofacial tissues from the perspective of developmental fate. Here, we review the fundamental biology and potential for future regeneration strategies of MSCs in oromaxillofacial research.


Asunto(s)
Odontología/tendencias , Células Madre Mesenquimatosas/citología , Regeneración , Diferenciación Celular , Pulpa Dental , Humanos
14.
Int J Mol Sci ; 20(3)2019 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-30759717

RESUMEN

Sjögren's syndrome (SjS) is an autoimmune disease that destroys the salivary glands and results in severe dry mouth. Mesenchymal stem cell (MSC) transplantation has been recently proposed as a promising therapy for restoring cells in multiple degenerative diseases. We have recently utilized advanced proteomics biochemical assays to identify the key molecules involved in the mesenchymal-epithelial transition (MET) of co-cultured mouse bone-marrow-derived MSCs mMSCs with primary salivary gland cells. Among the multiple transcription factors (TFs) that were differentially expressed, two major TFs were selected: muscle, intestine, and stomach expression-1 (MIST1) and transcription factor E2a (TCF3). These factors were assessed in the current study for their ability to drive the expression of acinar cell marker, alpha-salivary amylase 1 (AMY1), and ductal cell marker, cytokeratin19 (CK19), in vitro. Overexpression of MIST1-induced AMY1 expression while it had little effect on CK19 expression. In contrast, TCF3 induced neither of those cellular markers. Furthermore, we have identified that mMSCs express muscarinic-type 3 receptor (M3R) mainly in the cytoplasm and aquaporin 5 (AQP5) in the nucleus. While MIST1 did not alter M3R levels in mMSCs, a TCF3 overexpression downregulated M3R expressions in mMSCs. The mechanisms for such differential regulation of glandular markers by these TFs warrant further investigation.


Asunto(s)
Amilasas/metabolismo , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Células Madre Mesenquimatosas/metabolismo , Glándulas Salivales/metabolismo , Animales , Acuaporina 5/metabolismo , Biomarcadores/metabolismo , Células Cultivadas , Técnicas de Cocultivo/métodos , Regulación hacia Abajo/fisiología , Regulación de la Expresión Génica/fisiología , Humanos , Queratina-19/metabolismo , Ratones , Ratones Endogámicos C57BL , Proteómica/métodos , Síndrome de Sjögren/metabolismo , Factores de Transcripción/metabolismo
15.
Cell Physiol Biochem ; 46(1): 133-147, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29587276

RESUMEN

BACKGROUND/AIMS: Bone nonunion remains a challenge for orthopaedists. The technological advancements that have been made in precisely silencing target genes have provided promising methods to address this challenge. METHODS: We detected the expression levels of the bone morphogenetic protein (BMP) inhibitors Chordin, Gremlin and Noggin using realtime PCR in bone mesenchymal stem cells (BMSCs) isolated from patients with normal fracture healing and those with bone nonunion. Moreover, we detected the expression of Chordin, Gremlin and Noggin during the osteogenic differentiation of human BMSCs (hBMSCs) using real-time PCR and Western blot. We delivered Chordin siRNA to hBMSCs using a previously reported cationic polymer, polyspermine imidazole-4,5-imine (PSI), as a pH-responsive and non-cytotoxic transfection agent. The apoptosis and cellular uptake efficiency were analysed by flow cytometry. RESULTS: We identified Chordin as the most appropriate potential therapeutic target gene for enhancing the osteogenic differentiation of hBMSCs. Chordin knockdown rescued the osteogenic capacity of hBMSCs isolated from patients with bone nonunion. Highly efficient knockdown of Chordin was achieved in hBMSCs using PSI. Chordin knockdown promoted hBMSC osteogenesis and bone regeneration in vitro and in vivo. CONCLUSIONS: Our results suggest that Chordin is a potential target for improving osteogenesis and bone nonunion therapy and that responsive and non-toxic cationic polyimines such as PSI are therapeutically feasible carriers for the packaging and delivery of Chordin siRNA to hBMSCs.


Asunto(s)
Regeneración Ósea/fisiología , Glicoproteínas/metabolismo , Imidazoles/química , Péptidos y Proteínas de Señalización Intercelular/metabolismo , ARN Interferente Pequeño/metabolismo , Espermina/análogos & derivados , Células de la Médula Ósea/citología , Proteína Morfogenética Ósea 2/metabolismo , Proteína Morfogenética Ósea 4/metabolismo , Proteínas Portadoras/antagonistas & inhibidores , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Diferenciación Celular , Células Cultivadas , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Fracturas Óseas/patología , Glicoproteínas/antagonistas & inhibidores , Glicoproteínas/genética , Humanos , Concentración de Iones de Hidrógeno , Péptidos y Proteínas de Señalización Intercelular/química , Péptidos y Proteínas de Señalización Intercelular/genética , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Osteogénesis , Polietileneimina/química , Interferencia de ARN , ARN Interferente Pequeño/química , Proteína Smad1/metabolismo , Espermina/química
16.
Biochem Biophys Res Commun ; 497(4): 1011-1017, 2018 03 18.
Artículo en Inglés | MEDLINE | ID: mdl-29470980

RESUMEN

The objective was to investigate whether a graphene coating could improve the surface bioactivity of a cobalt-chromium-molybdenum-based alloy (CoCrMo). Graphene was produced by chemical vapor deposition and transferred to the surface of the CoCrMo alloy using an improved wet transfer approach. The morphology of the samples was observed, and the adhesion force and stabilization of graphene coating were analyzed by a nanoscratch test and ultrasonication test. In an in vitro study, the adhesion and proliferation of bone marrow mesenchymal stem cells (BMSCs) cultured on the samples were quantified via an Alamar Blue assay and cell counting kit-8 (CCK-8) assay. The results showed that it is feasible to apply graphene to modify the surface of a CoCrMo alloy, and the enhancement of the adhesion and proliferation of BMSCs was also shown in the present study. In conclusion, graphene exhibits considerable potential for enhancing the surface bioactivity of CoCrMo alloy.


Asunto(s)
Células de la Médula Ósea/citología , Materiales Biocompatibles Revestidos/química , Grafito/química , Células Madre Mesenquimatosas/citología , Vitalio/química , Células de la Médula Ósea/efectos de los fármacos , Adhesión Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Grafito/farmacología , Humanos , Células Madre Mesenquimatosas/efectos de los fármacos
17.
Biochem Biophys Res Commun ; 497(3): 876-882, 2018 03 11.
Artículo en Inglés | MEDLINE | ID: mdl-29477844

RESUMEN

Cleft lip and palate is the most common congenital anomaly in the orofacial region. Autogenous iliac bone graft, in general, has been employed for closing the bone defect at the alveolar cleft. However, such iliac bone graft provides patients with substantial surgical and psychological invasions. Consequently, development of a less invasive method has been highly anticipated. Stem cells from human exfoliated deciduous teeth (SHED) are a major candidate for playing a significant role in tissue engineering and regenerative medicine. The aim of this study was to elucidate the nature of bone regeneration by SHED as compared to that of human dental pulp stem cells (hDPSCs) and bone marrow mesenchymal stem cells (hBMSCs). The stems cells derived from pulp tissues and bone marrow were transplanted with a polylactic-coglycolic acid barrier membrane as a scaffold, for use in bone regeneration in an artificial bone defect of 4 mm in diameter in the calvaria of immunodeficient mice. Three-dimensional analysis using micro CT and histological evaluation were performed. Degree of bone regeneration with SHED relative to the bone defect was almost equivalent to that with hDPSCs and hBMSCs 12 weeks after transplantation. The ratio of new bone formation relative to the pre-created bone defect was not significantly different among groups with SHED, hDPSCs and hBMSCs. In addition, as a result of histological evaluation, SHED produced the largest osteoid and widely distributed collagen fibers compared to hDPSCs and hBMSCs groups. Thus, SHED transplantation exerted bone regeneration ability sufficient for the repair of bone defect. The present study has demonstrated that SHED is one of the best candidate as a cell source for the reconstruction of alveolar cleft due to the bone regeneration ability with less surgical invasion.


Asunto(s)
Regeneración Ósea , Pulpa Dental/citología , Células Madre Mesenquimatosas/citología , Trasplante de Células Madre , Diente Primario/citología , Proceso Alveolar/patología , Proceso Alveolar/fisiología , Diferenciación Celular , Células Cultivadas , Humanos , Trasplante de Células Madre Mesenquimatosas , Procedimientos de Cirugía Plástica , Medicina Regenerativa , Andamios del Tejido/química , Diente Primario/trasplante
18.
Osteoarthritis Cartilage ; 25(9): 1551-1562, 2017 09.
Artículo en Inglés | MEDLINE | ID: mdl-28532603

RESUMEN

OBJECTIVES: The present goal was to explore whether matrix replenishment is the primary requirement for osteoarthritic (OA) cartilage. METHODS: Cells isolated from the superficial and deep zone cartilage of a pig temporomandibular joint (TMJ) were exposed to fluid flow shear stress (FFSS). Differences in matrix production and cellular differentiation were detected. Unilateral anterior crossbite (UAC) was applied to C57BL/6J female mice. Green fluorescent protein-labeled exogenous bone marrow stromal cells (GFP-BMSCs) were injected weekly into TMJs, starting from 3 weeks of UAC stimulation and continuing for 4-, 8- and 12-weeks. Another GFP-BMSCs injection UAC group stopped receiving injections for 4-weeks after 8-weeks of injections. Assessments were focused on morphological alterations in UAC mouse TMJ cartilage, the expression levels of DAP3, an anoikis marker, CD163, a scavenger receptor family member, and ki67, a proliferation indicator. RESULTS: FFSS down-regulated type-II collagen expression but stimulated terminal differentiation in cells isolated from deep zone cartilage. It down-regulated aggrecan expression but up-regulated type I collagen in cells isolated from both superficial and deep zones. UAC caused matrix loss and anoikis and enhanced scavenging activity in deep zone chondrocytes without affecting cell proliferation. Superficial fibrillation was obvious in the late stage. Weekly injections of BMSCs largely restored these changes. The implanted BMSCs expressed a high level of CD163 protein but did not show remarkable cell proliferation. Terminating the supply of exogenous BMSCs reversed the restorative effects. CONCLUSIONS: Scavenging the degraded matrix and replenishing the fibrosis-developmental matrix are the primary requirements for the repair of OA cartilage.


Asunto(s)
Artritis Experimental/terapia , Matriz Ósea/patología , Trasplante de Células Madre Mesenquimatosas/métodos , Osteoartritis/terapia , Articulación Temporomandibular/patología , Animales , Artritis Experimental/metabolismo , Artritis Experimental/patología , Matriz Ósea/metabolismo , Cartílago Articular/metabolismo , Cartílago Articular/patología , Muerte Celular/fisiología , Diferenciación Celular/fisiología , Condrocitos/metabolismo , Colágeno Tipo II/metabolismo , Femenino , Maloclusión/metabolismo , Maloclusión/patología , Cóndilo Mandibular/metabolismo , Cóndilo Mandibular/patología , Células Madre Mesenquimatosas/fisiología , Ratones Endogámicos C57BL , Osteoartritis/metabolismo , Osteoartritis/patología , Fagocitosis/fisiología , Estrés Mecánico , Sus scrofa , Articulación Temporomandibular/metabolismo
19.
Odontology ; 105(4): 392-397, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27848099

RESUMEN

The major goal of dental pulp tissue engineering is to enable the healing of inflamed tissue or to replace necrotic pulp tissue with newly formed dental pulp tissue. Here, we report a protocol for pulp tissue engineering in vivo in pulpotomized rat teeth using constructs of rat bone marrow mesenchymal stem cells, preformed biodegradable scaffolds, and hydrogel. The constructs were implanted into pulpotomized pulp chambers for 3, 7, or 14 days. At 3 days, cells were located mainly along the preformed scaffolds. At 7 days, pulp tissue regeneration was observed in almost the entire implanted region. At 14 days, pulp tissue regeneration further progressed throughout the implanted region. In immunohistochemistry, at 3 days, a number of small and round macrophages immunoreactive to CD68 were predominantly distributed around the scaffolds. The density of CD68+ macrophages decreased until 14 days. On the other hand, nestin-expressing odontoblast-like cells beneath the dentin at the border of implanted region increased until 14 days. Quantitative gene expression analysis revealed that odontoblast differentiation marker dentin sialophosphoprotein mRNA in the implanted region gradually increased until 14 days. Together, the results suggested that regeneration of dental pulp tissue had occurred. Thus, our study provides a novel experimental rat model of dental pulp regeneration.


Asunto(s)
Pulpa Dental/citología , Células Madre Mesenquimatosas/citología , Ingeniería de Tejidos/métodos , Animales , Diferenciación Celular , Células Cultivadas , Cavidad Pulpar , Femenino , Hidrogeles/farmacología , Diente Molar , Pulpotomía , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Andamios del Tejido
20.
J Clin Periodontol ; 42(12): 1143-51, 2015 12.
Artículo en Inglés | MEDLINE | ID: mdl-26565741

RESUMEN

AIM: The aim of this study was to evaluate effective bone regeneration using an autologous serum scaffold (alone or seeded with autologous bone marrow-mesenchymal stem cells, BM-MSCs), when implanted in a 30 mm length segmental mandibular defect in sheep. MATERIALS AND METHODS: The bone defect was filled either with serum scaffold alone (control group; n = 5) or combined with BM-MSCs (experimental group; n = 10). Bone regeneration was determined at 12 (T12; 2 control sheep and 4 experimental sheep) and 32 weeks (T32; 3 control and 6 experimental sheep), as measured by computed and microcomputed tomography and histological examination. RESULTS: Two sheep of the Experimental group died after surgery. While complete bone union in the control group was only observed at T32, it was observed both at T12 (1/4 sheep) and T32 (3/4 sheep) in the experimental group. When properties/characteristics of new bone where compared, a better bone quality, similar to native bone, was observed in the scaffold combined with BM-MSCs. CONCLUSIONS: Based on these results, we conclude that the serum scaffold can promote efficient repair of large bone defects, but the combination with BM-MSCs accelerates this process, increasing significantly the amount and quality of bone formed.


Asunto(s)
Mandíbula , Animales , Células de la Médula Ósea , Células Madre Mesenquimatosas , Proyectos Piloto , Ovinos , Ingeniería de Tejidos , Andamios del Tejido , Microtomografía por Rayos X
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