Comparative immunohistochemucal evaluation of osteogenesis in large canine mandibular defect using adipose derived stem cells

Abstract

Introduction: Large mandibular defects are a major problem for oral and maxillofacial surgeons. Cell therapy as an alternative may speed up bone regeneration. The aim of this study was to evaluate bone regeneration in large mandibular defect in dogs using allogeneic undifferentiated fat stem cells on foam gel as cell carrier.

Method: In this basic experimental study with ethical code IR.TBZMED.VCR.REC.1400.114 with 6 dogs were examined. Tissue engineering phase includes: sampling of adult canine adipose tissue, isolation of canine adipose tissue stem cells (ASC), culture of ASCs in Dulbcco's Modified Eagle's Medium (DMEM) containing penicillin and streptomycin, identification of canine adipocytes, transplantation of ASCs To gelatin. In the next step, six dogs were randomly divided into three groups (group one titanium plate placement, group two titanium plate placement and foam gel, group three titanium plate placement and foam gel and stem cell). All three groups underwent surgery to create the same 3 cm bone defect and were reconstructed with titanium plates and screws. After 6 months, all groups were examined by immunohistochemistry to evaluate the degree of ossification at the defective site.

Results: Prior to bone grafting, the nature of stem cells was confirmed by flow cytometry and positive expression of CD105 and CD44 cell markers with 71.9% and 89.3%, respectively, and no expression of CD45 cell marker with 2.2%. The results of all three methods of H&E, IHC and Alizarin red staining showed more osteogenesis in the stem cell group than the control group and Scoffeld group

Conclution: Immunohistochemical results show that in large bone defects, the use of fat stem cells on the foam gel as cell carriers has a significant effect on bone formation.