Ground bone or cartilage particles are demineralized by extraction with a
low ionic strength buffer such as 20 mM HEPES containing a chelating agent
and protease inhibitors, then extracted with an acidic solution such as
0.3M citric acid, pH 4.0, containing protease inhibitors. The extracted
material generally contains less than 2% by weight phosphate and less than
100 mM calcium. The phosphate content can be further reduced by treatment
of the matrix with acid phosphatase, which removes residual organic
phosphate. The material is useful in a method of treatment of vesicouretal
reflux and other disorders where a bulking agent is effective is
correcting the defect.
