Mesenchymal stem cell-conditioned media: A novel alternative of stem cell therapy for quality wound healing.

Mesenchymal stem cells-conditioned media (MSCs-CM) contains several growth factors and cytokines, thus may be used as a better alternative to stem cell therapy, which needs to be elucidated. The present study was conducted to evaluate the therapeutic potential of caprine, canine, and guinea pig bone marrow-derived MSCs-CM in excision wound healing in a guinea pig model. MSCs were obtained from bone marrow, expanded ex vivo and characterized as per ISCT criteria. CM was collected assayed by western blot to ascertain the presence of important secretory biomolecules. Quantitative estimation by enzyme-linked immunosorbent assay was done for a vascular epidermal growth factor (VEGF) and interleukin-6 (IL-6) in caprine MSCs-CM and optimum time for collection of CM was decided as 72 hr. CM from all the species was lyophilized by freeze-drying method. Full-thickness (2 × 2 cm2 ) excision skin wounds were created in guinea pigs (six animals in each group) and respective lyophilized CM mixed with laminin gel was applied topically at weekly interval. On Day 28, histopathological examinations of healed skin were done by hemotoxylin and eosin staining. MSCs were found to secrete important growth factors and cytokines (i.e., VEGF, transforming growth factor-ß1, fibroblast growth factor-2, insulin-like growth factor-1, stem cell factor, and IL-6) as demonstrated by immunohistochemistry and western blot assay. It was found that allogenic and xenogenic application of CM significantly improved quality wound healing with minimal scar formation. Thus, MSCs-CM can be used allogenically as well as xenogenically for quality wound healing.

Expression pattern of GLUT 1, 5, 8 and citrate synthase transcripts in buffalo (Bubalus bubalis) preimplantation embryos produced in vitro and derived in vivo.

In vitro-produced (IVP) embryos are reported to be developmentally lesser competent than in vivo-derived (IVD) embryos and supposed to differ in the expression of genes related with glucose metabolism. So, the present study was conducted to analyse the expression pattern of GLUT 1, 5, 8 and citrate synthase (CS) in oocytes and embryos produced in vivo or in vitro in buffalo. IVD embryos were obtained from 18 superovulated buffaloes. IVP embryos were obtained from slaughterhouse-derived oocytes subsequently subjected to in vitro fertilization and culture. Total RNA was isolated from different stages of oocytes (immature and in vitro matured) and embryos (8-16 cell to blastocysts of IVP embryos and morula to blastocysts of IVD embryos). Results demonstrated that the expression of GLUT1, GLUT 8 increased from 8 to 16 cells to blastocyst and was significantly (p < .05) higher in IVP embryos. Expression of both genes was (p < .05) higher in IVD than in IVP blastocysts; though GLUT5 transcripts were not detected at 8- to 16-cell stage IVP embryos, significantly (p < .05) higher transcripts were found at morula and blastocyst stages irrespective of embryo source with significantly (p < .05) higher expression in IVD embryos compared to IVP embryos. No significant difference was observed in citrate synthase expression in embryos at morula stage irrespective of the embryo source while significantly (p < .05) higher transcript level was observed at blastocyst stage with no difference between in vivo and in vitro embryos. It can be concluded that expression of GLUTs and CS is upregulated with progression of embryonic stage and expression is higher in in vivo embryos than in vitro counter parts; thus, it can be said that in vivo-produced embryos are metabolically superior to in vitro embryos.

Comparative evaluation of fracture healing potential of differentiated and undifferentiated guinea pig and canine bone marrow-derived mesenchymal stem cells in a guinea pig model.

BACKGROUND AND AIM: This work was conducted to compare the therapeutic potential of undifferentiated and osteogenic differentiated canine (xenogeneic) and guinea pig (allogeneic) BMSCs in fracture healing using guinea pig as a model. MATERIALS AND METHODS: A well-characterized homogenous population of third passage mesenchymal stem cells of bone marrow origin was used in all the experiments. MSCs from both the species, i.e., canine and guinea pigs, were differentiated and characterized. Expression of MHC I and II along with co-stimulatory molecules was assessed based on relative mRNA expression. The osteogenic differentiated and undifferentiated MSCs from both species were used for evaluating fracture healing in the guinea pig model. The healing potential was assessed based on radiographic, histopathology, and clinical observations. RESULTS: BMSCs from both species expressed MSC surface antigens and successfully differentiated to osteogenic, chondrogenic, and adipogenic lineages. The mRNA expression of class I and II MHC molecules in all the three lineages showed no significant (p > 0.05) differences after differentiating to adipogenic, chondrogenic, and osteogenic lineages. Radiographic and clinical examination revealed that MSCs therapy significantly improved bone fracture healing with a non-significant (p > 0.05) difference between differentiated and undifferentiated BMSCs. In addition, allogeneic MSCs therapy performed better than xenogeneic therapy. CONCLUSION: MSCs remained hypo immunogenic after differentiation and have comparable fracture healing potential though allogeneic MSCs have better therapeutic potential than xenogenic MSCs.