A Combination Histochemical and Autoradiographic Method for Analysis of Enzymatic and Proliferative Responses to Immunoreactive Osteoinducer
Abstract
Although bone matrix derivatives have been shown to induce new bond formation, little is known about the mechanism of matrix-induced osteogenesis. We produced antisera and monoclonal antibodies reactive with bovine bone morphogenetic protein (bBMP) and applied them to tissue sections alone and in combination with histochemical and autoradiographic methods. In this way, we studied the in vivo fate of immunologically defined components of the osteoinducer and analyzed their tissue distribution relative to proliferative and enzymatic responses associated with new bone formation. Two monoclonal antibodies, reactive with bBMP in tissue sections, revealed diffusion of the relevant determinant from implanted matrix particles and its incorporation by responding cells. A method for simultaneously demonstrating immunoreactive bBMP, DNA replication and alkaline phosphate activity in the responding cells was shown to result in effective differential labeling of these features in mildly fixed tissue sections. Application of this method with monoclonal antibodies specific for induction-associated determinants and with modifications to permit ultrastructural analyses may provide important information relevant to the mechanism of matrix-induced bone formation. Keywords: Osteogenic implants; Immunohistochemistry; Histochemistry.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 15, 1988
- Accession Number
- ADA199430
Entities
People
- James W. Mizgala
- Paul R. Burnett
Organizations
- United States Army Institute of Surgical Research