Bone Marrow-Derived Mesenchymal Stromal Cells Combined with Biodegradable Scaffold and Biological Therapy Using Growth Factor (Vegf), Hyperbaric Oxygen and Low-Intensity Pulsed Ultrasound to Improve Muscle Healing after Laceration Injury

Muscle injuries are frequently encountered in professional and recreational sports. The best treatment for these injuries has not yet been clearly defined. Our series researches have demonstrated that bone marrow-derived mesenchymal stromal cells (BMMSCs), hyperbaric oxygen (HBO) and low-intensity pulsed ultrasound (LIPUS) could improve muscle healing after injury (NSC91-2314-B182A-079, NSC93-2314-B-182A-027, NSC94-2314-B-182A-049, NSC95-2314-B-182A-084, and NSC96-2628-B-182A-085). However, the previous three biological therapies for muscle healing still had the functional limitation of recovery compared with normal muscle. Stem cells are undifferentiated cells capable of proliferation, self-renewal, production of a large number of differentiated progeny, and regeneration of tissues. The authors have identified a novel population of bone marrow-derived mesenchymal stromal cells from mice: potential for muscle regeneration (NSC96-2628-B-182A-085). Soft-tissue engineering to repair defective muscle, require a soft muscle conductive vehicle. The biodegradable scaffold, diblock copolymer, methoxy poly (ethylene glycol) poly(ε-capro-lactone) (MPEG–PCL) have been used as effective tools for targeted cell delivery. These in vivo gel-forming, degradable scaffolds exhibit temperature-dependent phase-change properties, converting from a solution to a gel at approximately body temperature. Thus, when injected into the body, the scaffold forms a gel-type scaffold, providing a matrix for the proliferation and differentiation of delivered cells without the need for additional surgery. Angiogenic processes that establish and maintain blood vessel networks are important for preventing muscle cell death and tissue necrosis. Enhanced angiogenesis may thus improve muscle function in ischemic tissue. Vascular endothelial cell growth factor (VEGF), HBO and LIPUS are potent mediators of angiogenesis In the current 3-year study, the authors design a series of researches to prove the hypothesis that the combination of BMMSCs and in vivo gel-forming scaffolds containing an angiogenesis-enhancing growth factor (VEGF: vascular endothelial cell growth factor), HBO or LIPUS are suitable for muscle tissue engineering. Specifically, we evaluated muscle regeneration, strength recovery and blood vessel formation in vivo. (1) First-year proposal: To focus on the combination effects in vitro of BMMSCs with biodegradable scaffold, diblock copolymer, methoxy poly (ethylene glycol) poly (ε-capro-lactone) (MPEG–PCL). (2) Second-year proposal: To focus on the relationship of angiogenesis-enhancing factors (VEGF, HBO or LIPUS) times, therapy dosage and mechanism on BMMSCs with MPEG–PCL in vitro. Global gene expression pattern will be studied by DNA microarray. Specific aims of this proposal includes :(1) to test the interaction of angiogenesis-enhancing factors on BMMSCs with MPEG–PCL. (2) to identify critical genes in BMMSCs differentiation under different therapy dosage. . (3) Third-year proposal: To focus on the in vivo study using our well-established muscle laceration injury model in mice. The authors will follow the in vitro results of previous 2 years researches. Evaluation by histology, immunohistochemical staining, RT-PCR, Western blot, DNA microarray and muscle physiological test, the effects and mechanism of BMMSCs with MPEG–PCL, VEGF, HBO and LIPUS on muscle healing after injury will be identified. All the information obtained from this proposal will be the fundamental keystones for future researches on muscle healing using tissue engineering for clinical application.

Project ID:PC10001-1413
External Project ID:NSC99-2314-B182A-098-MY3