Enhanced Healing of Rat Calvarial Bone Defects with Hypoxic Conditioned Medium from Mesenchymal Stem Cells through Increased Endogenous Stem Cell Migration via Regulation of ICAM-1 Targeted-microRNA-221

Abstract

The use of conditioned medium from mesenchymal stem cells may be a feasible approach for regeneration of bone defects through secretion of various components of mesenchymal stem cells such as cytokines, chemokines, and growth factors. Mesenchymal stem cells secrete and accumulate multiple factors in conditioned medium under specific physiological conditions. In this study, we investigated whether the conditioned medium collected under hypoxic condition could effectively influence bone regeneration through enhanced migration and adhesion of endogenous mesenchymal stem cells. Cell migration and adhesion abilities were increased through overexpression of intercellular adhesion molecule-1 in hypoxic conditioned medium treated group. Intercellular adhesion molecule-1 was upregulated by microRNA-221 in mesenchymal stem cells because microRNAs are key regulators of various biological functions via gene expression. To investigate the effects in vivo, evaluation of bone regeneration by computed tomography and histological assays revealed that osteogenesis was enhanced in the hypoxic conditioned medium group relative to the other groups. These results suggest that behavioral changes of endogenous mesenchymal stem cells through microRNA-221 targeted-intercellular adhesion molecule-1 expression under hypoxic conditions may be a potential treatment for patients with bone defects.

Conditioned medium of mesenchymal stem cells delays osteoarthritis progression in a rat model by protecting subchondral bone, maintaining matrix homeostasis, and enhancing autophagy

“…Hayami et al () verified that the bone resorption inhibitor could effectively protect articular cartilage in the rat model of OA. Then in , Chang et al observed the osteogenesis potent of MSCs–CM in healing bone defects. Hence, it is possible that the osteogenesis potent of CM maintained the balance between resorption and formation of subchondral bone, which further preserved the microarchitecture of subchondral bone.…”

Alterations in Cardiomyocyte Differentiation-Related Proteins in Rat Mesenchymal Stem Cells Exposed to Hypoxia

“…Primary rMSCs were isolated and cultured according to the optimized methods described in our previous studies [26,27]. Briefly, BM from the femurs and tibias of 4-week-old Sprague-Dawley male rats was flushed with phosphate-buffered saline (PBS; HyClone, Logan, UT, USA) containing 2% fetal bovine serum (FBS; HyClone), and mononuclear cells were isolated via density gradient centrifugation (Ficoll-Paque Plus; GE Healthcare, Sweden) for 30 min at 1,000 ×g.…”

“…Isolated BM-rMSCs from the femurs and tibias of Sprague-Dawley male rats were cultured according to the optimized methods described in our previous studies [26,27]. Isolated rMSCs were characterized based on the presence or absence of specific markers using flow cytometry; that is, they expressed CD90, CD105, and CD106, but not CD34, CD45, and CD14 (Fig.…”

Adipose Derived Stem Cells Conditioned Media in Combination with Bioceramic-Collagen Scaffolds Improved Calvarial Bone Healing in Hypothyroid Rats

“…Inukai et al showed a positive effect of multiple cytokines contained in MSC-CM on alveolar bone and cementum regeneration (32). Also, Chang et al reported that paracrine factors secreted from MSC under hypoxic condition could enhance healing of the calvarial defect in rats through increased endogenous stem cell migration via regulation of ICAM-1 (intercellular adhesion molecule-1) targeted microRNA-221 (37). However, most previous in vivo studies were performed on normal animal models and no investigation has been done to date using ADMSC-CM for bone healing in a hypothyroid animal model.…”