A novel method to improve the osteogenesis capacity of hUCMSCs with dual‐directional pre‐induction under screened co‐culture conditions

Abstract

Objectives Mesenchymal stem cells (MSCs) based therapy for bone regeneration has been regarded as a promising method in the clinic. However, hBMSCs with invasive harvesting process and undesirable proliferation rate hinder the extensive usage. HUCMSCs of easier access and excellent performances provide an alternative for the fabrication of tissue‐engineered bone construct. Evidence suggested the osteogenesis ability of hUCMSCs was weaker than that of hBMSCs. To address this issue, a co‐culture strategy of osteogenically and angiogenically induced hUCMSCs has been proposed since thorough vascularization facilitates the blood‐borne nutrition and oxygen to transport in the scaffold, synergistically expediting the process of ossification. Materials and methods Herein, we used osteogenic‐ and angiogenic‐differentiated hUCMSCs for co‐culture in screened culture medium to elevate the osteogenic capacity with in vitro studies and finally coupled with 3D TCP scaffold to repair rat's critical‐sized calvarial bone defect. By dual‐directional induction, hUCMSCs could differentiate into osteoblasts and endothelial cells, respectively. To optimize the co‐culture condition, gradient ratios of dual‐directional differentiated hUCMSCs co‐cultured under different medium were studied to determine the appropriate condition. Results It revealed that the osteogenic‐ and angiogenic‐induced hUCMSCs mixed with the ratio of 3:1 co‐cultured in the mixed medium of osteogenic induction medium to endothelial cell induction medium of 3:1 possessed more mineralization nodules. Similarly, ALP and osteogenesis/angiogenesis‐related genes expressions were relatively higher. Further evidence of bone defect repair with 3D printed TCP of 3:1 group exhibited better restoration outcomes. Conclusions Our work demonstrated a favourable and convenient approach of dual‐directional differentiated hUCMSCs co‐culture to improve the osteogenesis, establishing a novel way to fabricate tissue‐engineered bone graft with 3D TCP for large bone defect augmentation.

Fabrication of a bio-instructive scaffold conferred with a favorable microenvironment allowing for superior implant osseointegration and accelerated in situ vascularized bone regeneration via type H vessel formation

“…The local microenvironment in diseased or traumatized bone is usually altered or compromised, manifesting as the impaired formation of blood vessels and new bone due to a disruption of coupled angiogenesis and osteogenesis [ 30 , 46 ]. The reciprocal relationship of neovascularization and bone regeneration is regulated by complex crosstalk between endothelial cells and osteoprogenitors [ 9 , 30 , [47] , [48] , [49] , [50] , [51] , [52] , [53] , [54] , [55] , [56] ]. In response to chemoattractants, ECs and osteoprogenitors are recruited by direct migration from their niches or defect edges and then proliferate and differentiate into vessels and bone [ 57 ].…”

Clinically relevant preclinical animal models for testing novel cranio‐maxillofacial bone 3D‐printed biomaterials

“…The CMF defect models discussed in this review are the calvarial, mandibular and orbital floor defect models. In most cases, a trephine 119–152 is used to create cylindrical defects in the calvaria and the mandible. Other bone cutting devices for cylindrical defects include a circular knife, 153 a biopsy punch, 154 a drill 126,155–159 and rongeurs 160 .…”

Osteogenically Induced Human Umbilical Cord Mesenchymal Stem Cell-derived Small Extracellular Vesicles Carrying Distinct microRNA Signature Improve Osteoinductivity of Tissue-engineered Bone

“…The primary antibodies included TSG101 (1:2000; abcam, USA), Alix (1:2000; abcam, USA), and Calreticulin (1:2000; abcam, USA). The specific procedures were performed as previously reported (22).…”

“…Fabrication and observation of 3D printed TCP scaffold: The scaffolds were fabricated as previously described (22). The scaffold with a diameter of 5 mm, a thickness of 1 mm and a porosity of 200 μm were eventually obtained.…”

“…Besides, they also showed excellent restorative outcome when combined with 3D printed tricalcium phosphate (TCP) scaffold to repair rat calvarial defects (19). It has also been widely reported that the secreted factors of hUCMSCs can promote the osteogenic differentiation of MSCs and repair calvarial defects (20).…”