Alveolar Repair Using Cancellous Bone and Beta Tricalcium Phosphate Seeded With Adipose-Derived Stem Cell.

INTRODUCTION: Adipose-derived stem cells (ADSCs) have been subject of several studies due to their abundance, ease of preparation, and application in bone regeneration. We aim to compare effectiveness of alveolar reconstruction utilizing human cancellous freeze-dried graft (HCG) and beta tricalcium phosphate (BTP), both seeded with human ADSC (hADSC) and autologous bone graft (ABG). MATERIAL AND METHODS: A 5 × 5 mm alveolar defect in 36 male Wistar rats were treated using: ABG (C), HCG-hADSC (H1), and BTP-hADSC (H2). At 1 and 8 weeks after surgery, runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osterix (OSX), and bone morphogenetic protein 2 (BMP2; g/mL) were quantified using immunohistochemistry, while bone tissue volume (BV, mm3), bone tissue volume fraction (BF, percentage), and trabecular thickness of bone (TT, mm) were assessed using micro-computed tomography (CT). RESULTS: One week after surgery, H2 was higher in RUNX2, OSX, ALP, and BMP2 than C (P < .05). Only RUNX2 and OSX were found to be higher in H1 than C, while ALP and BMP2 were higher in H2 than H1. Micro-CT revealed that H2 had a higher TT than C and C had a higher TT than H1 (P < .05). Eight weeks after surgery, both H2 and H1 was higher in RUNX2, OSX, ALP, and BMP2 than C (P < .05). RUNX2 and BMP2 were found to be higher in H1 than H2. Micro-CT revealed that H2 had higher BV and TT than C and H1 (P < .05). CONCLUSIONS: Exogenous hADSC strengthened the effectiveness of HCG and BTP to accelerate osteogenesis, osteoconduction, and osteoinduction. The latter was the most successful in bone formation, followed by HCG and ABG.

The Characteristics of Demineralized Dentin Material Sponge as Guided Bone Regeneration Based on the FTIR and SEM-EDX Tests.

OBJECTIVE: The objective of this study was to determine the characteristics of demineralized dentin material sponge (DDMS). MATERIAL AND METHODS: An observational study was conducted on DDMS and BPCM. Fourier transform infrared (FTIR) test was performed to determine the characterizations of the materials. Scanning electron microscope-electron dispersive X-ray spectroscopy (SEM-EDX) test was performed to observe the elements contained in the materials. RESULTS: The infrared spectrum of the DDMS and BPCM functional groups showed the same pattern in each variation, and no significant differences were found. According to SEM analysis, the cavities that make up the membrane were spotted on the surface. Besides, according to the SEM-EDX analysis, DDMS contained chlorine, carbon, and calcium, while BPCM contained carbon, oxygen, and sulfur. CONCLUSION: DDMS has the potential to be a biomaterial for bone tissue engineering in terms of the characteristics. DDMS had a structure that almost resembles BPCM as seen from the results of the FTIR graph between DDMS and BPCM. The morphological structure of the two materials in the SEM test appeared to have porosity with various sizes.

Extracellular Vesicles from Stem and Progenitor Cells for Cell-Free Regenerative Therapy.

Cell-based regenerative therapies involving stem or progenitor cells are considered as possible therapeutic modalities to treat non-communicable and degenerative diseases. Recently, regenerative outcomes of cell-based therapies have been linked to paracrine factors and extracellular vesicles [EVs] released by the transplanted cells rather than the transplanted cells themselves. EVs contain a cargo that includes microRNAs [miRNAs], mRNAs, as well as proteins. Their role in mediating intercellular communication has been acknowledged in several studies. However, the regenerative potential of the miRNAs, mRNAs, and proteins that are present in EVs is a matter of ongoing scientific debate. In this review, we discuss EVs as an alternative to stem cell-based therapy to treat some of the non-communicable and degenerative diseases. Moreover, we also propose that pre-treatment of the cells could help to produce EVs enriched with particular miRNAs, mRNAs, and/or proteins that could support the successful regeneration of a targeted organ.

In vitro Cell Proliferation Assay of Demineralized Dentin Material Membrane in Osteoblastic MC3T3-E1 Cells.

AIM: Demineralized dentin material membrane (DDMM) is a novel bioresorbable guided bone regeneration (GBR) which is derived from the demineralization process of bovine dentin. This material/process could be an alternative to resolve musculoskeletal dysfunction that harms the quality of human life. PURPOSE: To evaluate the cytotoxic effect of DDMM as GBR membrane on MC3T3-E1 osteoblast cell line. METHODS: Cytotoxic effect was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Osteoblast MC3T3-E1 cell culture was used as a parameter of cell viability after reacting with GBR materials. The absorbance values were examined at each treatment to determine the percentage of cell viability. There were four groups created in the present study: two treatment groups and two control groups. The treatment groups consisted of a DDMM group and a bovine pericardium collagen membrane (BPCM) group. The control groups comprised a group containing cell culture medium as a negative control group and another positive control group that contained cell cultures. RESULTS: The results revealed no significant difference in MC3T3-E1 cell viability between the treatment and control groups (p < 0.05). Moreover, as observed in the DDMM group, there was an increase in the number of osteoblast cells. CONCLUSION: DDMM is a suitable alternative biomaterial for GBR as it is non-cytotoxic and could potentially increase the rate of repair of craniofacial defects.

Role of the CXCR4-SDF1-HMGB1 pathway in the directional migration of cells and regeneration of affected organs.

In recent years, several studies have reported positive outcomes of cell-based therapies despite insufficient engraftment of transplanted cells. These findings have created a huge interest in the regenerative potential of paracrine factors released from transplanted stem or progenitor cells. Interestingly, this notion has also led scientists to question the role of proteins in the secretome produced by cells, tissues or organisms under certain conditions or at a particular time of regenerative therapy. Further studies have revealed that the secretomes derived from different cell types contain paracrine factors that could help to prevent apoptosis and induce proliferation of cells residing within the tissues of affected organs. This could also facilitate the migration of immune, progenitor and stem cells within the body to the site of inflammation. Of these different paracrine factors present within the secretome, researchers have given proper consideration to stromal cell-derived factor-1 (SDF1) that plays a vital role in tissue-specific migration of the cells needed for regeneration. Recently researchers recognized that SDF1 could facilitate site-specific migration of cells by regulating SDF1-CXCR4 and/or HMGB1-SDF1-CXCR4 pathways which is vital for tissue regeneration. Hence in this study, we have attempted to describe the role of different types of cells within the body in facilitating regeneration while emphasizing the HMGB1-SDF1-CXCR4 pathway that orchestrates the migration of cells to the site where regeneration is needed.

The role of salivary sIgA as protection for dental caries activity in Indonesian children.

PURPOSE: The aim of this study is to asses the relationship between the level of sIgA and dental caries experience in healthy children who are 6- to 9-years-old from Indonesia. The case-control study is conducted to determine the protective role of salivary secretory immunoglobulin A (sIgA) levels in the stimulated whole saliva of dental caries-active and caries-free children. METHODS: This research was done by stimulating the whole saliva which had been collected from 6- to 9-years-old children with the index def-t≥3 of 30 children as the caries-active children group and 30 children with def-t<3 as the low caries-active children group. Saliva samples were collected in sterile vials between 10 am-12 pm due to the circadian rhythm, which is at least one hour after last meal. 1,5 ml of collected salivary sample was centrifuged, then the supernatants was transferred to other tube and stored immediately to the laboratory at a temperature of -20 °C. The estimation of sIgA concentration was done by using ELISA. The differences in the level of sIgA between the two groups with caries were analyzed using the t-test afterward. RESULTS: The total salivary concentration of sIgA was statistically significantly higher in the low caries-active children group than in the caries-active children group. CONCLUSION: The total salivary concentration of sIgA was statistically and significantly higher in the low caries-active children group than caries-active children Group. There is a negative correlation between sIgA level and dental caries activity of 6 to 9-years-old children.

Detection of Candida albicans biofilm proteins induced by glucose, lactose, soy protein, and iron.

BACKGROUND: Oral candidiasis is one of the most common fungal infections, which attack the mucosa of the oral cavity. These lesions are mostly caused by the fungal species Candida albicans. Candida albicans is included in the normal oral microorganisms that are opportunistic pathogens, and its presence is quite large, which can reach 75% of the total oral fungal population. Research on specific proteins of Candida biofilm can be an alternative to early prevention of oral infections such as Oral Candidiasis. This biofilm protein can be used as a reference in making kits to detect the presence of microbes that cause infectious diseases. The purpose of this study was to determine molecular weight of Candida albicans biofilm protein induced by 5% glucose, 5% lactose, soy protein, and 5% iron. MATERIAL AND METHODS: This experimental laboratory study used SDS-PAGE electrophoresis to determine the molecular weight of Candida albicans biofilm proteins induced by glucose 5%, lactose 5%, soy protein, and iron 5%. RESULTS: Biofilm induced by 5% glucose shows four protein bands: 71,6 kDa; 56,1 kDa; 49,7 kDa; and 41 kDa. Biofilm induced by 5% lactose shows seven protein bands: 71 kDa; 61,2 kDa; 57,7 kDa; 55,3 kDa; 48,9 kDa; 39,5 kDa; and 29,8 kDa. Biofilm induced by soy protein shows one protein band: 49,4 kDa. Biofilm induced by 5% iron shows one protein band: 51,1 kDa. CONCLUSIONS: Candida albicans biofilm induced by 5% glucose has four protein band candidates, 5% lactose has seven candidates of protein band, and soy protein and 5% iron each has a candidate of protein band, which can be used as a target for the detection of oral Candidiasis. Key words:Biofilm protein, Candida albicans, molecular weight, oral candidiasis.

Demineralized Freeze-Dried Bovine Cortical Bone: Its Potential for Guided Bone Regeneration Membrane.

BACKGROUND: Bovine pericardium collagen membrane (BPCM) had been widely used in guided bone regeneration (GBR) whose manufacturing process usually required chemical cross-linking to prolong its biodegradation. However, cross-linking of collagen fibrils was associated with poorer tissue integration and delayed vascular invasion. OBJECTIVE: This study evaluated the potential of bovine cortical bone collagen membrane for GBR by evaluating its antigenicity potential, cytotoxicity, immune and tissue response, and biodegradation behaviors. MATERIAL AND METHODS: Antigenicity potential of demineralized freeze-dried bovine cortical bone membrane (DFDBCBM) was done with histology-based anticellularity evaluation, while cytotoxicity was analyzed using MTT Assay. Evaluation of immune response, tissue response, and biodegradation was done by randomly implanting DFDBCBM and BPCM in rat's subcutaneous dorsum. Samples were collected at 2, 5, and 7 days and 7, 14, 21, and 28 days for biocompatibility and tissue response-biodegradation study, respectively. RESULT: DFDBCBM, histologically, showed no retained cells; however, it showed some level of in vitro cytotoxicity. In vivo study exhibited increased immune response to DFDBCBM in early healing phase; however, normal tissue response and degradation rate were observed up to 4 weeks after DFDBCBM implantation. CONCLUSION: Demineralized freeze-dried bovine cortical bone membrane showed potential for clinical application; however, it needs to be optimized in its biocompatibility to fulfill all requirements for GBR membrane.