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1.
Br J Neurosurg ; 37(3): 485-494, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32648779

RESUMEN

BACKGROUND: Ultrasonography (US) provides real-time information on structures within the skull during neurosurgical operations. Superb microvascular imaging (SMI) is the latest imaging technique for detecting very low-velocity flow with minimal motion artifacts, and we have reported on this technique for intraoperative US monitoring. We combined SMI with administration of contrast agent to obtain detailed information during neurosurgical operations. MATERIALS AND METHODS: Twenty patients diagnosed with brain tumor (10 meningiomas, 5 glioblastomas, 2 hemangioblastomas, 1 schwannoma, 1 malignant lymphoma, 1 brain abscess) underwent neurosurgery under US with SMI and contrast agent techniques. Vessel density and appearance time following contrast administration were analyzed. RESULTS: Flow in numerous vessels was not visualized by SMI alone, but appeared following injection of contrast agent in all cases. Flow in tumors was drastically enhanced by contrast agent in schwannoma, hemangioblastoma and meningioma, compared to normal brain tissue. Flows in the dilated and bent vessels of glioblastoma were also enhanced, although flow in hypoechoic lymphoma remained inconspicuous. The characteristics of tumor vessels were clearly visualized and tumor borders were demonstrated by the difference between tumor flow and brain flow, by the increased tumor vessel density and decreased appearance time of contrast agent compared to normal brain vessels. CONCLUSIONS: The combination of SMI and contrast agent techniques for intraoperative US monitoring could provide innovative flow images of tumor and normal brain. The neurosurgeon obtains information about tumor flow and tumor borderline before tumor resection.


Asunto(s)
Neoplasias Encefálicas , Neoplasias Meníngeas , Meningioma , Neurilemoma , Humanos , Medios de Contraste , Ultrasonografía/métodos , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/cirugía , Meningioma/diagnóstico por imagen , Meningioma/cirugía , Neoplasias Meníngeas/diagnóstico por imagen , Neoplasias Meníngeas/cirugía , Neurilemoma/diagnóstico por imagen , Neurilemoma/cirugía
2.
Biotechnol Lett ; 43(6): 1241-1251, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33768381

RESUMEN

PURPOSE: McCarey-Kaufman's (MK) medium and Optisol-GS medium are the most commonly employed media for human donor corneal preservation. In this study, we evaluated the preservation efficacy of discarded human donor corneas using a Thermo-reversible gelation polymer (TGP) added to these two media. METHODS: Thirteen human corneal buttons collected from deceased donors, which were otherwise discarded due to low endothelial cell density (ECD) were used. They were stored in four groups: MK medium, MK medium with TGP, Optisol-GS and Optisol-GS with TGP at 4 °C for 96 h. Slit lamp examination and specular microscopy were performed. Corneal limbal tissues from these corneas were then cultured using explant methodology one with and the other without TGP scaffold, for 21 days. RESULTS: MK + TGP and Optisol-GS + TGP preserved corneas better than without TGP, which was observed by maintenance of ECD which was significantly higher in Optisol-GS + TGP than MK + TGP (p-value = 0.000478) and corneal thickness remaining the same for 96 h. Viable corneal epithelial cells could be grown from the corneas stored only in MK + TGP and Optisol-GS + TGP. During culture, the TGP scaffold helped maintain the native epithelial phenotype and progenitor/stem cell growth was confirmed by RT-PCR characterization. CONCLUSION: TGP reconstituted with MK and Optisol-GS media yields better preservation of human corneal buttons in terms of relatively higher ECD maintenance and better in vitro culture outcome of corneal limbal tissue. This method has the potential to become a standard donor corneal transportation-preservation methodology and it can also be extended to other tissue or organ transportation upon further validation.


Asunto(s)
Medios de Cultivo/química , Endotelio Corneal/citología , Conservación de Tejido/métodos , Adulto , Anciano , Anciano de 80 o más Años , Cadáver , Sulfatos de Condroitina/química , Mezclas Complejas/química , Dextranos/química , Femenino , Gentamicinas/química , Humanos , Masculino , Persona de Mediana Edad , Compuestos Orgánicos/química , Microscopía con Lámpara de Hendidura
3.
Thromb J ; 18: 27, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33082714

RESUMEN

Direct endothelial injury by viruses and dysregulation of clotting mechanisms due to cytokine storm are the major precipitating factors of mortality in COVID-19; both are attributed to a fundamental dysregulation of the immune system. While immune dysregulation can be attributed to several factors, the risk of associated thrombogenic disruption varies across individuals. This variation depends on several factors, such as comorbidities, including diabetes, hypertension, and cardiovascular diseases. When considering ethnic variations, the vulnerability of Caucasians, African Americans and Hispanics needs to be addressed before arriving at strategies to handle thromboembolic complications, which have been identified in recent reports as the leading causes of mortality in COVID-19. Although evaluation of D-dimer and prothrombin during admission is considered to predict prognosis and mortality, there are no preventive or prophylactic strategies before hospital admission. Herein, we present our perspectives on the effect of regular supplementation with the biological response modifier beta glucan based on its relevance to immune modulation. This effect is of paramount importance in decreasing the development of severe COVID-19 and reducing mortality against the background of coagulopathy, especially in vulnerable populations.

4.
Cureus ; 15(5): e39010, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-37323332

RESUMEN

We describe the first case of a 62-year-old female symptomatic patient with multiple comorbidities presenting with coronal limb malalignment due to scoliosis and osteoarthritis who underwent a combined total hip arthroplasty and biplane opening wedge osteotomy of the distal femur in one procedure. It is essential to realize that in patients who present with multiple comorbidities, combining different established procedures should be considered as a therapeutic option. We detail the operative procedure, preoperative considerations, and postoperative rehabilitation. By reviewing the literature on operative techniques, we highlight how our findings can be applied to similar cases with multiple comorbidities. Our report underscores the importance of considering combined procedures as a viable therapeutic option for patients with complex medical histories.

5.
Cureus ; 15(10): e46395, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37927684

RESUMEN

Background Postoperative surgical site infections (SSIs) are a significant complication of surgical procedures, leading to increased morbidity, prolonged hospital stays, and substantial healthcare costs; however, the use of drain tip cultures to diagnose SSIs in patients is controversial. The objective of this study was to evaluate the efficacy of drain tip cultures for the prediction of postoperative SSIs in patients recovering from hip arthroplasty. Methodology The data were collected from 1204 patients who underwent hip arthroplasty procedures over 15 years, and statistical analysis was performed to evaluate the diagnostic value of drain tip culture in determining surgical site infection. We also used these data to evaluate whether preexisting conditions such as hypertension or diabetes affected the probability of a patient getting an SSI. Results Drain tip cultures were positive in 12 of 1,112 cases of primary hip arthroplasty, but only one of these 12 patients was ultimately diagnosed with an SSI (sensitivity, 12.5%; specificity, 99.0%; p = 0.0834). Results from postoperative drain tip cultures performed in patients undergoing revision arthroplasty included two false positives and three false negatives; interestingly, no true positives were detected in any of the revision arthroplasty cases we evaluated (sensitivity, 0%; specificity, 97.8%; p = 0.9355). Conclusion Our results indicate that drain tip cultures have no statistically significant predictive value for the diagnosis of postoperative SSIs and thus should not be used as a primary diagnostic or predictive tool for SSIs. We recommend exploring other diagnostic tools for the postoperative diagnosis of SSIs. Standardized guidelines should therefore be established to improve the predictive value of the different methods.

6.
Curr Eye Res ; 48(6): 564-572, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-36852699

RESUMEN

PURPOSE: Corneal limbal stem cell (LSC) transplantation has been reported as a potential approach to treat the damaged corneal epithelium. Scaffolds such as human amniotic membrane (hAM) are commonly employed for the in vitro culture and as a carrier during in vivo transplantation. However, they carry the risk of biological contamination and donor to donor variability. To overcome these disadvantages, we herein report the capabilities of a synthetic thermoreversible gelation polymer (TGP) scaffold to serve as an encapsulation support during LSC transplantation and to enable engraftment for corneal regeneration. METHODS: Sixteen discarded human corneas were used to isolate the corneal epithelium which was cultured in TGP and hAM. The cell proliferation and characteristics between TGP and hAM culture methods were evaluated by microscopic observation, 3H Thymidine incorporation assay, immunoperoxidase and immunofluorescence staining. RESULTS: The 3H Thymidine assay's results showed that TGP allowed human-donor cornea-derived LSCs to proliferate well in vitro, compared to hAM and the cells encapsulated in TGP and transplanted ex vivo onto a human cadaver donor cornea denuded of its epithelium, migrated on the ocular surface, and proliferated to form a continuous layer in 25 days. Immunoperoxidase and Immunofluorescence staining of TGP-cultured cells were positive for LSC markers (p63, ABCG2, Connexin 43 and Integrin ß), proving that the TGP helps to preserve the limbal cells' stemness. CONCLUSION: TGP is found to be a multipurpose scaffold for (i) in vitro culture, (ii) ex vivo encapsulation, and in vivo transplantation (iii), enabling engraftment of LSCs in this study, with potentials to extend its application in cell-based therapies in several regenerative medicine approaches.


Asunto(s)
Trasplante de Córnea , Epitelio Corneal , Limbo de la Córnea , Humanos , Córnea , Epitelio Corneal/metabolismo , Células Cultivadas , Proliferación Celular
7.
J Orthop ; 29: 60-64, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35145328

RESUMEN

BACKGROUND: Chondrocytes are used in cell-based therapies such as autologous chondrocyte implantation (ACI) and matrix-associated cartilage implantation (MACI). To transport the cartilage tissue to the laboratory for in vitro culturing, phosphate-buffered saline (PBS), Euro-Collins solution (ECS) and Dulbecco's Modified Eagle's Medium (DMEM) are commonly employed at 4-8 °C. METHODS: In this study, eight samples of human cartilage biopsy tissues from elderly patients with severe osteoarthritis undergoing arthroscopy, which would otherwise have been discarded, were used. The cartilage tissue samples were compared to assess the cell yield between two transportation groups: i) a thermo-reversible gelation polymer (TGP) based method without cool preservation (∼25 °C) and ii) ECS transport at 4 °C. These samples were subjected to in vitro culture in a two-dimensional (2D) monolayer for two weeks and subsequently in a three-dimensional (3D) TGP scaffold for six weeks. RESULTS: The cell count obtained from the tissues transported in TGP was higher (0.2 million cells) than those transported in ECS (0.08 million cells) both after initial processing and after in vitro culturing for 2 weeks in 2D (18 million cells compared with 10 million cells). In addition, mRNA quantification demonstrated significantly higher expression of Col2a1 and SOX-9 in 3D-TGP cultured cells and lower expression of COL1a1 in RT-PCR, characteristic of the hyaline cartilage phenotype, than in 2D culture. CONCLUSION: This study confirms that the TGP cocktail is suitable for both the transport of human cartilage tissue and for in vitro culturing to yield better-quality cells for use in regenerative therapies.

8.
Biopreserv Biobank ; 20(1): 97-103, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-34962137

RESUMEN

Introduction: The transportation of tissues from hospitals to clinical laboratories for cell therapy is an essential component of regenerative medicine. Previously, we used laboratory-cultured mucosal cells from buccal epithelium expanded and encapsulated using a scaffold-hybrid approach to the urethral stricture (BEES-HAUS) procedure. In this study, to improve the outcomes, we compared the thermoreversible gelation polymer (TGP) transportation procedure with conventional culture methods, and reported its advantages. Methods: Human buccal mucosal tissues in Phase I of the study were transported in Euro-Collins solution (ECS) and the cells obtained were cultured in two-dimensional (2D) Dulbecco's modified Eagle's medium (DMEM), CnT-Prime epithelial 2D differentiation medium (CnT-PR), and a three-dimensional (3D)-TGP scaffold. In Phase II, tissues were transported in a TGP cocktail and the ECS. The cells were cultured in 2D-DMEM and 3D-TGP, quantified, and characterized by immunohistochemistry. Results: The cells in 3D-TGP culture maintained epithelial morphology in a better manner compared with 2D-DMEM, in which they developed fibroblast-like morphology. The TGP-transported cells grew rapidly. Immunohistochemical analysis results for AE1/AE3, EGFR, integrin-ß1, p63, and p75 were intensely positive in 3D-TGP. Conclusion: The TGP-based cocktail used in human buccal tissue transportation yielded cells with better morphology maintenance. The TGP scaffold provides an optimal in vitro environment wherein epithelial cells better maintain their native phenotype compared to those cultured through conventional methods. These results suggest using TGP for the transportation and culture of human buccal tissues for clinical applications. In addition, the use of a TGP-based cocktail for the transport of other tissues for regenerative medicine applications is worth further analysis.


Asunto(s)
Ingeniería de Tejidos , Estrechez Uretral , Células Cultivadas , Células Epiteliales , Humanos , Polímeros , Ingeniería de Tejidos/métodos
9.
J Clin Neurosci ; 89: 329-335, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-34119288

RESUMEN

Intraoperative ultrasound during transsphenoidal surgery (TSS) for pituitary tumors has been reported. In reports of endonasal ultrasound (US), Doppler US vessel images were informative and effective in endoscopic TSS. We performed endoscopic US imaging with high flow mode, which is a novel technology, to visualize small vessels during endonasal endoscopic TSS. Six patients (five with pituitary adenomas and one with Rathoke's cleft cyst) underwent endoscopic US-assisted TSS. A small endoscopic US probe (Olympus, BF-UC260FW; diameter, 6.9 mm) was inserted transsphenoidally to the sellar floor and into the sella turcica, and endoscopic US monitoring was performed. By rotating the endoscopic US probe, the internal carotid artery, anterior cerebral artery, middle cerebral artery, various small vessels, optic nerve, and residual tumor were clearly visualized on the endoscopic US images. Real-time animated vessel images around the tumor could be generated when needed during TSS. The tumors were removed without leakage of cerebrospinal fluid in the six patients, and their visual acuity was restored. Endoscopic US with high flow mode can visualize not only main cerebral arteries but also intracranial small vessels on B-mode US images. Pituitary tumors were clearly recognized and removed safely and precisely by monitoring the cerebral artery and its small branches as landmarks.


Asunto(s)
Adenoma/cirugía , Cirugía Endoscópica por Orificios Naturales/métodos , Neoplasias Hipofisarias/cirugía , Ultrasonografía/métodos , Adenoma/diagnóstico por imagen , Adulto , Pérdida de Líquido Cefalorraquídeo/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Cirugía Endoscópica por Orificios Naturales/efectos adversos , Nariz/cirugía , Neoplasias Hipofisarias/diagnóstico por imagen , Complicaciones Posoperatorias/epidemiología , Silla Turca/efectos de los fármacos , Silla Turca/cirugía
10.
Orthop Res Rev ; 13: 63-71, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34017204

RESUMEN

We describe the case of a 76-year-old Asian female patient who presented with severe pain and a valgus deformity of the right knee. Her past medical history included neurofibromatosis, which resulted in a severe anterior slope of the right knee, limb shortening, and congenital pseudarthrosis. She was diagnosed with severe anterolateral osteoarthritis and eburnation of the right knee that was treated surgically with a lateral unicompartmental knee arthroplasty (UKA). Bone and cartilage fragments were removed from the joint space and a UKA implant (Zimmer®) with dimensions of 29 mm × 50 mm was inserted. Perioperative imaging revealed that the procedure resulted in the correction of the valgus deformity. Pain was reduced and the patient was able to walk for 10 meters with support during the immediate postoperative period. One week post-surgery, the patient sustained an oblique tibial fracture that extended from the medial edge of the implant to the medial slope of the proximal tibia. This complication may have been attributed to large implant size or sagittal overcutting. The fracture was treated surgically with a rotated anterolateral locking plate (A.L.P.S®) inserted into the distal tibia. The patient was capable of ambulation at full weight load at two months after the second procedure. It is critical to recognize that there are no standard protocols that can be used to guide the treatment of neurofibromatosis-induced osteoarthritis. The specific preoperative condition of the individual patient plays a large role in determining the appropriate treatment option. In this case, the availability of custom-fitted UKA implants might have improved outlook, we understand that these devices are costly and may not be available at all hospitals. However, we strongly believe that the "gold standard" in these cases is patient-specific treatment that addresses the issues of the highest concern using the resources that are available at that time.

11.
Life Sci ; 278: 119553, 2021 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-33932445

RESUMEN

AIMS: We have evaluated the potential of a three-dimensional (3D) thermoreversible gelation polymer (TGP) matrix in enhancing miRNA 140 expression (a biomarker correlating with homeostasis and cartilage regeneration) during the in vitro expansion of osteoarthritis (OA)-affected human chondrocytes. MATERIALS AND METHODS: OA-chondrocytes were cultured in two-dimensional (2D) monolayer followed by culture in 3D-TGP. miRNA 140 expression levels in cell culture supernatant followed by expression in the cell lysate of both 2D and 3D-TGP cultures were analyzed. KEY FINDINGS: The expression of miRNA 140 in cell culture supernatant from the 3D-TGP group was 0.001 to 0.002% that in 2D culture supernatant while in the cell lysate, miRNA 140 expression in the 3D-TGP was nearly 30-fold higher than that of 2D group. SIGNIFICANCE: The 3D-TGP matrix allows enhanced expression of miRNA 140 in OA-affected human chondrocytes in vitro which after necessary validations can be applied in clinical transplantation to significantly improve the outcome.


Asunto(s)
Condrocitos/patología , MicroARNs/genética , Osteoartritis/genética , Regulación hacia Arriba , Técnicas de Cultivo de Célula , Células Cultivadas , Condrocitos/metabolismo , Humanos , Osteoartritis/patología , Polímeros/química , Andamios del Tejido/química
12.
Knee ; 29: 365-373, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33690017

RESUMEN

BACKGROUND: Chondrocyte transplantation to address cartilage damage is an established solution. Because hyaluronic acid (HA) is an essential component for homeostasis of the cartilage, in order to arrive at methodologies to utilize its advantages in cell-based therapies, we compared the HA retention capability of a thermoreversible gelation polymer scaffold-based environment (3D-TGP) with conventional in vitro cell culture methodologies. METHODS: Chondrocytes derived from osteoarthritis-affected knee joint cartilage of elderly patients were used and accomplished in three phases. In Phase I, the levels of HA secreted by chondrocytes were measured in culture supernatant. In Phase II, retention capacity of externally added HA was quantified indirectly by measuring the HA released in culture supernatant, and in Phase III, the expression of CD44 on cells was analysed by immunohistochemistry. RESULTS: In Phase I, the average HA in the 3D supernatant was 3% that of 2D. In phase II, 80% of externally added HA was detected in the 2D on day 7, while in 3D-TGP, only 0.1% was released until day 21. In Phase III, 2D yielded individual cells that started degenerating from the third week; in 3D-TGP cells grew for a longer duration, formed a tissue-like architecture with extracellular matrix with significantly intense staining of CD44 than 2D. CONCLUSION: The capability of the 3D-TGP culture environment to retain HA and support chondrocytes to grow with a tissue-like architecture expressing higher HA content is considered advantageous as it serves as an in vitro culture platform that enables tissue engineering of cartilage tissue with native hyaline phenotype and higher HA expression. The in vitro environment being conducive, based on this data, we also recommend that the TGP be tried as an encapsulation material in clinical studies of chondrocyte implantation for optimal clinical outcome.


Asunto(s)
Cartílago Articular/citología , Condrocitos/metabolismo , Ácido Hialurónico/metabolismo , Osteoartritis de la Rodilla/metabolismo , Andamios del Tejido , Anciano , Células Cultivadas , Humanos , Receptores de Hialuranos/metabolismo , Inmunohistoquímica
13.
Sci Rep ; 11(1): 14059, 2021 07 07.
Artículo en Inglés | MEDLINE | ID: mdl-34234261

RESUMEN

Regenerative medicine applications require cells that are not inflicted with senescence after in vitro culture for an optimal in vivo outcome. Methods to overcome replicative senescence include genomic modifications which have their own disadvantages. We have evaluated a three-dimensional (3D) thermo-reversible gelation polymer (TGP) matrix environment for its capabilities to reverse cellular senescence. The expression of senescence-associated beta-galactosidase (SA-ßgal) by human chondrocytes from osteoarthritis-affected cartilage tissue, grown in a conventional two-dimensional (2D) monolayer culture versus in 3D-TGP were compared. In 2D, the cells de-differentiated into fibroblasts, expressed higher SA-ßgal and started degenerating at 25 days. SA-ßgal levels decreased when the chondrocytes were transferred from the 2D to the 3D-TGP culture, with cells exhibiting a tissue-like growth until 42-45 days. Other senescence associated markers such as p16INK4a and p21 were also expressed only in 2D cultured cells but not in 3D-TGP tissue engineered cartilage. This is a first-of-its-kind report of a chemically synthesized and reproducible in vitro environment yielding an advantageous reversal of aging of human chondrocytes without any genomic modifications. The method is worth consideration as an optimal method for growing cells for regenerative medicine applications.


Asunto(s)
Biopolímeros , Senescencia Celular/genética , Condrocitos/metabolismo , Regulación de la Expresión Génica , Ingeniería de Tejidos , Andamios del Tejido , beta-Galactosidasa/genética , Biomarcadores , Técnicas de Cultivo de Célula , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Ingeniería de Tejidos/métodos , beta-Galactosidasa/metabolismo
14.
J Orthop ; 23: 138-141, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33510554

RESUMEN

OBJECTIVE: We evaluated the expression of stem/progenitor biomarkers in osteoarthritic tissue derived chondrocytes cultured using a three-dimensional (3D) thermo-reversible gelation polymer (TGP). METHODS: The chondrocytes from discarded biopsy tissues obtained from human elderly patients with osteoarthritis were cultured using the 3D-TGP up to six weeks. RESULTS: The chondrocytes grew in a tissue-like manner, without de-differentiation into fibroblasts, and the cells thus tissue-engineered were proven positive for CD49e, OCT4, CD-105 and STRO-1 by immunohistochemistry. CONCLUSION: This study establishes the efficacy of this 3D-TGP platform for clinically useable in-vitro tissue-engineered cartilage for improvising the clinical outcome of cell therapy for cartilage repair.

15.
Curr Stem Cell Res Ther ; 16(4): 385-399, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33121414

RESUMEN

BACKGROUND: Cell-based therapies represent one of the definitive treatment approaches to SCI which, to become a routine clinical application, is marred by several known unknowns. The Bone Marrow Mononuclear Cells (BMMNCs) and Mesenchymal Stem Cells (MSCs) represent the most clinically applied cell types for SCI in humans, with safety established, and to an extent, efficacy reported. METHODS: In this review, we have analysed the clinical studies performed using BMMNC and MSC for complete SCI separately, and the potential for applying those cells in combination. We have also analysed those factors whose outcome in animal studies of SCI could be evaluated in depth but the clinical outcome cannot be evaluated intrinsically owing to practical difficulties. CONCLUSION: A combination of these two cell types, BMMNC and MSC, has been proven to be advantageous than applying them separately. Therefore, a thorough evaluation including the rationale and potential implications of applying these two therapies has been presented here, and we hypothesize that such a combination is likely to improvise the outcome of a wholesome approach to spinal cord regeneration after SCI.


Asunto(s)
Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , Traumatismos de la Médula Espinal , Animales , Médula Ósea , Células de la Médula Ósea , Humanos , Traumatismos de la Médula Espinal/terapia
16.
Regen Ther ; 18: 127-132, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34189194

RESUMEN

BACKGROUND: A pilot study reported an autologous buccal mucosal cell transplant in humans through the trans-urethral route using the buccal epithelium expanded and encapsulated in scaffold-hybrid approach to urethral stricture (BEES-HAUS), a minimally invasive approach to treat urethral stricture. Although successful outcomes were achieved in that study, for further validation, it is essential to prove that the transplanted buccal epithelium was engrafted over the urothelium through histological examination of the urethra, harvested post-transplant, which is infeasible in humans. Herein, we report the successful creation of an animal model of urethral stricture and the engraftment of epithelial cells derived from autologous buccal mucosal tissue, encapsulated in a thermo-reversible gelation polymer (TGP) scaffold, transplanted by trans-urethral route. METHODS: An animal model of urethral stricture was created in Japanese white male rabbits using electro-coagulation. Buccal tissue was harvested from the rabbits and subjected to enzyme digestion, followed by 5-7 days of in vitro culture in conventional two-dimensional (2D) culture and in a 3D platform of thermo-reversible gelation polymer (3D-TGP) culture. The cells harvested from the groups were mixed and encapsulated and transplanted with TGP, by transurethral catheterization. Fourteen days later, the urethra was harvested and subjected to histological examination. The buccal biopsy tissue, cells after digestion and cells post-culture were also subjected to histological examination. Urethrogram and endoscopy images were recorded at different time points. RESULTS: The stricture was successfully created, with the coagulated area markedly stenosed. Histological staining of the cells after in vitro processing showed that the cells grew with native epithelial and rounded cell morphology in 3D-TGP while they differentiated into fibroblast like-cells in 2D culture. Histological staining of the urethral tissue after transplantation revealed the engraftment of the transplanted buccal mucosal cells, with stratified squamous epithelium over the specialized stratified urothelium in the urethrotomy site. CONCLUSION: We used histology to prove the successful engraftment of TGP-encapsulated buccal mucosal epithelial cells in an animal model of urethral injury with healing of the injured tissue. The model of urethral stricture and cell therapy, using a transurethral approach, recapitulates the previously reported BEES-HAUS approach and lays the foundation for larger multi-centric translational clinical studies.

17.
Tokai J Exp Clin Med ; 46(1): 1-6, 2021 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-33835468

RESUMEN

BACKGROUND: Buccal mucosal epithelial cells show promising application for various regenerative medicine approaches. In this study, we examined the feasibility of culturing rabbit and human buccal mucosal epithelial cells in a novel thermoreversible gelation polymer (TGP) scaffold, without feeder layers or other foreign proteins. METHODS & RESULTS: The results of this 28-day in vitro culture, u sing the conventional technique (2D) and TGP (3D) showed that the epithelial cell morphology could be maintained only in the TGP group while cells in the 2D group de-differentiated to fibroblast morphology in both human and rabbit samples. CK3 expression, a marker for epithelial differentiation was higher in 3D-TGP cultured cells than 2D. CONCLUSION: TGP based in vitro cell culture is a prospective methodology to culture buccal mucosal epithelial cells efficiently without using foreign biological components for tissue engineering applications.


Asunto(s)
Técnicas de Cultivo de Célula , Tratamiento Basado en Trasplante de Células y Tejidos , Células Epiteliales/fisiología , Mucosa Bucal/citología , Polímeros , Ingeniería de Tejidos/métodos , Andamios del Tejido , Animales , Diferenciación Celular/genética , Células Cultivadas , Células Epiteliales/metabolismo , Estudios de Factibilidad , Fibroblastos , Expresión Génica , Humanos , Queratina-3/genética , Queratina-3/metabolismo , Conejos , Factores de Tiempo
18.
Orthop Res Rev ; 12: 189-193, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33364860

RESUMEN

The present case report describes a pediatric patient who presented with flexor digitorum profundus (FDP) entrapment after a forearm fracture. The patient was diagnosed with a Bado type I Monteggia fracture. The fracture was reduced using closed reduction under fluoroscopy followed by Kirschner's wire fixation. This case is unique because the FDP was found entrapped in the fracture site 2 weeks post-operation and was managed using conservative therapy. FDP entrapment is typically managed by surgical intervention, and there have been no previous reports of conservative management. The FDP was released using passive extension of the index finger under general anesthesia, and no irreversible damage to the tendon or muscle was found. This case report demonstrates the potential for conservative therapy in the management of FDP entrapment after forearm fractures.

19.
Arch Acad Emerg Med ; 8(1): e54, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32440665

RESUMEN

COronaVIrus Disease 2019 (COVID-19) is an on-going pandemic attributed to a novel virus named SARS-CoV-2. Comparing the statistics of incidence and death rates between nations reveals that there is discrepancy amongst countries in these regards, even between countries that share borders. We herein present information from the literature indicating how cross-protection against COVID-19 conferred by the encephalitis vaccine could be the reason for lower fatality rate in the countries where immunization against encephalitis is widespread or included in national programs. This may pave the way for arriving at efficient prevention strategies as well as vaccine development.

20.
Regen Ther ; 14: 234-237, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32435676

RESUMEN

Autologous chondrocytes in vitro expanded, are used as tools of regenerative therapies for cartilage injuries. However, inability to maintain the hyaline phenotype both in vitro and post in vivo transplantation, remains one of the major hurdles for long term efficacy under clinical settings. We have reported earlier, hyaline phenotype maintenance of both human and rabbit chondrocytes for a long duration both in vitro when cultured conditions using a Thermo-reversible Gelation Polymer (TGP) scaffold-based methodology and in vivo post-transplantation animal model of cartilage damage. Having intrigued by such encouraging outcome, we in this study, analysed the similar TGP culture environment whether would be able to allow in vitro expansion of severe osteoarthritis affected cartilage tissue from elderly patients and evaluated the cells using lectin microarray characterization for pluripotency. Cartilage tissue were obtained from patients (n = 7; age: 60-85 years) undergoing total knee arthroplasty for severe osteoarthritis. Chondrocytes were isolated and cultured in two groups: i. conventional culture without scaffold (2D) and ii. using a TGP scaffold-based culture (3D) up to 18 weeks. In addition to earlier reported findings such as maintenance of hyaline phenotype having been confirmed in this study as well, surface glycoprotein analysis by lectin microarray demonstrated that the α1-2 Fuc recognition lectin (UEA-1) (marker reported in literature for pluripotent stem cells) was found to be more highly expressed in 3D culture compared to 2D culture and even increased over time in 3D culture. We have developed an environment where osteoarthritis affected chondrocytes from the elderly could be cultured up to 18 weeks in vitro using TGP scaffold which express pluripotent cell associated surface glycoproteins compared to the conventional methodology.

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