Dedifferentiated fat cells (DFATs) isolated from mature adipocytes have a multilineage differentiation capacity similar to mesenchymal stem cells and are considered as promising source of cells for tissue engineering. Bone morphogenetic protein 9 (BMP9) and low-intensity pulsed ultrasound (LIPUS) have been reported to stimulate bone formation both in vitro and in vivo. However, the combined effect of BMP9 and LIPUS on osteoblastic differentiation of DFATs has not been studied. After preparing DFATs from mature adipose tissue from rats, DFATs were treated with different doses of BMP9 and/or LIPUS. The effects on osteoblastic differentiation were assessed by changes in alkaline phosphatase (ALP) activity, mineralization/calcium deposition, and expression of bone related genes; Runx2, osterix, osteopontin. No significant differences for ALP activity, mineralization deposition, as well as expression for bone related genes were observed by LIPUS treatment alone while treatment with BMP9 induced osteoblastic differentiation of DFATs in a dose dependent manner. Further, co-treatment with BMP9 and LIPUS significantly increased osteoblastic differentiation of DFATs compared to those treated with BMP9 alone. In addition, upregulation for BMP9-receptor genes was observed by LIPUS treatment. Indomethacin, an inhibitor of prostaglandin synthesis, significantly inhibited the synergistic effect of BMP9 and LIPUS co-stimulation on osteoblastic differentiation of DFATs. LIPUS promotes BMP9 induced osteoblastic differentiation of DFATs in vitro and prostaglandins may be involved in this mechanism.

PURPOSE: Furcation involvement in the molars is difficult to treat, and has been recognized as a risk factor for tooth loss. Although periodontal regenerative therapies, including guided tissue regeneration and various types of bone grafts, have been applied to furcation defects, the effects of these treatments are limited, especially in large class III furcation defects. The purpose of this pilot study was to investigate the effect of reciprocal autologous root transplantation on periodontal wound healing and regeneration in class III furcation defects in dogs. METHODS: Furcation defects (7 mm wide and 6 mm high) were surgically created after root separation of the unilateral third and fourth premolars in 4 dogs. Eight furcation defects were randomized to receive either reciprocal autologous root transplantation (test) or no further treatment (control). In the test group, the mesial and distal roots were transplanted into the distal and mesial extraction sockets, respectively. The animals were sacrificed 10 weeks after surgery for histologic evaluation. RESULTS: The healing pattern in the control group was characterized by extensive collapse of the flap and limited periodontal regeneration. New bone formation in the test group (3.56±0.57 mm) was significantly greater than in the control group (0.62±0.21 mm). Dense collagen fibers inserting into the residual cementum on the transplanted root surfaces were observed in the test group. Slight ankylosis was observed in 2 of the 4 specimens in the test group on the mesiodistal sides where the root-planed surfaces faced the existing bone. Root resorption (RR) was detected in both the control and test groups. CONCLUSIONS: Within the limits of this study, it can be concluded that reciprocal autologous root transplantation was effective for bone regeneration in class III furcation defects in dogs. However, further studies are required to standardize the approach in order to prevent unwanted RR prior to clinical application.
Animals ; Ankylosis ; Bicuspid ; Bone Regeneration ; Collagen ; Dental Cementum ; Dogs ; Furcation Defects* ; Guided Tissue Regeneration ; Models, Animal ; Molar ; Osteogenesis ; Periodontal Diseases ; Periodontal Ligament ; Pilot Projects ; Regeneration ; Risk Factors ; Root Resorption ; Tooth Loss ; Transplants ; Wound Healing* ; Wounds and Injuries*