United States Patent [19]
Brockbank
US005131850A
[ii] Patent Number: [45] Date of Patent:
[54] METHOD FOR CRYOPRESERVING MUSCULOSKELETAL TISSUES
[75] Inventor: Kelvin G. M. Brockbank, Marietta, Ga.
[73] Assignee: Cryolife, Inc., Marietta, Ga.
[21] Appl. No.: 431,153
[22] Filed: Nov. 3,1989
[51] Int. CI.' A01N 1/02
[52] U.S. CI 435/1; 62/62;
435/2
[58] Field of Search 62/62; 514/54; 435/1,
435/2; 436/8-18; 623/16
[56] References Cited
U.S. PATENT DOCUMENTS
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3,444,039 5/1969 Rajamannan 435/2
4,486,416 12/1984 Soil et al 514/54
4,800,429 11/1989 Stone 623/16
4,959,319 9/1990 Skelnik et al 435/240.2
OTHER PUBLICATIONS
Vasseur et al., Clinical Orthopedics and Related Re-
search, 219, pp. 268-277 (1987).
Frank et al., "Viability of Ligaments After Freezing:
An Experimental Study in a Rabbit Model", Journal of
Orthopaedic Research, 6:95-102 (1988).
Arnoczky et al., "The Effect of Cryopreservation on
Canine Menisci: A Biochemical, Morphologic, and
Biomechanical Evaluation", J. of Orthopaedic Re-
search, (1988).
"Meniscal Transplantation in Goats: An Experimental
Study", E. M. Keating, M.D., Center for Hip and Knee
Surgery, 34th Annual Meeting, Orthopaedic Research
Society, Feb. 1-4, 1988.
Schachar et al., "Investigations of Low-Temperature
Storage of Articular Cartilage for Transplantation",
Clinical Orthopaedics, Jul. 1986, vol. 208, pp. 146-150.
Fu et al., "The Science of Anterior Cruciate Ligament
Implants—1989", American Academy of Orthopaedic
Surgeons 56th Annual Meeting, (1989).
Nikolaou et al., "Anterior Cruciate Ligament Allograft
Transplantation" Long-Term Function, Histology,
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348-360, The American Journal of Sports Medicine,
vol. 14, No. 5 (1986).
Primary Examiner—Sam Rosen
Attorney, Agent, or Firm—Cushman, Darby & Cushman
[57] ABSTRACT
Disclosed herein is a method for cryopreserving musculoskeletal tissues, such as ligaments, tendons and cartilage, by placing such tissue in contact with a composition containing a cryopreserving agent comprising a cell-penetrating organic solute, which is preferably dimethylsulfoxide, and a glycosaminoglycan, which is preferably chondroitin sulphate, in an amount sufficient to cryopreserve the musculoskeletal tissue. The addition of a glycosaminoglycan to a cryopreserving agent comprising a cell-penetrating organic solute permits a broad range of cooling rates to be employed, rather than the very narrow ranges which are employed using a cryopreserving agent comprising a cell-penetrating organic solvent without the glycosaminoglycan. Also disclosed are a freezing schedule designed to maximize retention of tissue cell viability and biomechanical properties during and after the freezing process, and a thawing schedule which maximizes cell viability.
9 Claims, 3 Drawing Sheets
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