Introduction Experimental heterotopic bone formation in the canine urinary bladder has been observed for more than seventy years without revealing the origin of the osteoinductive signals. In 1931, Huggins demonstrated bone formation in a fascial transplant to the urinary bladder. Through an elaborate set of experiments, it was found that proliferating canine transitional epithelial cells from the urinary system act as a source of osteoinduction.
Urist performed a similar series of experiments in guinea pigs as Huggins did in his canine model. After two weeks, mesenchymal cells condensed against the columnar epithelium and membranous bone with haversian systems and marrow began to form juxtapose the basement membrane. At no time was cartilage formation noted, only direct membranous bone formation. They also demonstrated the expression of BMP’s in migrating epithelium and suggested that BMP is the osteoinductive factor in heterotopic bone formation.
Method This study was approved by Institutional Animal Ethics Committee. Six dogs underwent a mid-line laparotomy incision followed by mobilisation of a right sided myoperitioneal vascularised flap based on an inferior epigastric artery pedicle. A sagittal cystotomy is made in the dome of the bladder and the vascularised flap was sutured in place with acryl absorbable, continuous suture. The animals were sacrificed at 6 weeks. The bladder samples were removed and assessed by histology and immunohistochemistry. Sections were incubated with optimal dilution of primary antibody for type I collagen, type III collagen, alkaline phosphatase (ALP), bone morphogenetic protein (BMP)-2 and –4, osteocalcin (OCN), osteopontin (OPN), bone sialoprotein (BSP).
Results The mechanism for bone formation induced by the epithelial-mesenchymal cell interactions is not clear. We were able to demonstrate mature lamellar bone formation 6 weeks after transplanting a portion of the abdominal smooth muscle into the bladder wall. The bone formed immediately adjacent to the proliferating transitional uroepithelium, a prerequisite for bone formation in Huggins’ model. Here we report evidence of cartilage formation and therefore endochondral ossification as well as membranous bone formation. This is very similar histologically to the process of endochondral ossification at the growth plate in the growing skeleton. We propose a mechanism for the expression of BMP by epithelial cells.
Discussion This study demonstrates transitional epithelium induced formation of chondrocytes and osteoblasts in muscle tissue. The sequential expression of bone matrix proteins was related to cell proliferation, differentiation and formation of endochondral and membranous bone. Further information regarding the molecular mechanism of bone formation induced by epithelial-mesenchymal cell interactions will improve understanding of cell differentiation during osteogenesis.
