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A synthetic polypentapeptide of elastin for initiating calcification

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Summary

A polypentapeptide (PPP) of tropoelastin having a repeating amino acid sequence of (Val-Pro-Gly-Val-Gly)n was evaluated for its potential to initiatein vivo calcification and to enhance bone formation in nonhealing calvarial wounds (8.0 mm) in 396 adult Walter Reed rats. There were four configurations of the PPP (molecular weight range of 50–100K dalton) consisting of 1-dry PPP; 2-co-acervate PPP; 3-gamma irradiated, cross-linked PPP; 4-calcified, gamma irradiated, cross-linked PPP. These four iterations plus a control group of animals constituted the five treatment classes that were evaluated at days 1, 3, 7, 21, 42, and 147. Seventy two rats were used for each treatment and 36 rats for the control. Following euthanatization, specimens were placed into 70% ethanol, embedded in polymethyl methacrylate, sectioned at 3.5 micrometers, and alternating sections were stained with Masson-Goldner trichrome and von Kossa stains.

Histomorphometric analysis was accomplished using a Zeiss Universal microscope (250×) and Videoplan Image Analysis System to evaluate five random histologic fields from margin to margin of the craniotomy. Trabecular bony volume and area of calcification islands were quantitated. A Student'st test for unpaired data to determine treatment differences (within the same temporal groups) revealed that there was no significant difference between treatments and control for trabecular bony volume; however, there was a significant difference between experimentals and control for calcification islands (P<0.05) such that calcifications islands for the experimentals were greater than the control. There was not a significant difference between experimental treatments. These data provide the firstin vivo demonstration of the validity of the neutral site binding/charge neutralization mechanism of calcification. Although the synthetic peptide did support calcification, in no group was new bone formed in the craniotomies.

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Authors and Affiliations

  1. Laboratory of Molecular Biophysics, University of Alabama at Birmingham, School of Medicine, Birmingham, Alabama, U.S.A.

    M. M. Long, K. U. Prasad & D. W. Urry

  2. United States Army Institute of Dental Research, 20307-5300, Washington, D.C.

    J. O. Hollinger, J. P. Schmitz & Ruth Yaskovich

Authors
  1. J. O. Hollinger
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  2. J. P. Schmitz
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  3. Ruth Yaskovich
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  4. M. M. Long
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  5. K. U. Prasad
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  6. D. W. Urry
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Additional information

Commercial materials and equipment are identified in this report to specify the investigative procedure. Such identification does not imply recommendation or endorsement, or that the materials and equipment are necessarily the best available for the purpose. Furthermore, the opinions expressed here are those of the authors and are not to be construed as those of the Army Medical Department. In conducting research described in this report, the investigators adhered to the “Guide for the Care and Use of Laboratory Animals” as promulgated by the Committee on the Revision of the Guide for Laboratory Animal Facilities and Care of the Institute of Laboratory Animal Resources National Research Council.

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Hollinger, J.O., Schmitz, J.P., Yaskovich, R. et al. A synthetic polypentapeptide of elastin for initiating calcification. Calcif Tissue Int 42, 231–236 (1988). https://doi.org/10.1007/BF02553748

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  • DOI: https://doi.org/10.1007/BF02553748

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