Growth of osteoid osteoma transplanted into athymic nude mice.

An osteoid osteoma, excised from the neck of the femur of a 23-year-old man, was cut into four 1.5 mm3 fragments and immediately transplanted into muscle pouches in athymic nude mice. One fragment was devitalized by lyophilization before implantation.

Degradation of bone matrix morphogenetic activity by pulverization.

The yield of new bone from implants of pulverized demineralized whole matrix and bone matrix gelatin declines as the particle size decreases in diameter below 125 microns. The corresponding increase in surface area and mechanically-induced free radicals is associated with an increase in solubility of bone matrix proteins. These changes in physiocochemical properties and the concomitant reduction in bone yield suggest that prolonged pulverization denatures a bone morphogenetic protein (BMP).

Solubilized and insolubilized bone morphogenetic protein.

A bone morphogenetic protein (BMP) obtained in solution by digestion of demineralized rabbit cortical bone matrix with bacterial collagenase retains its biologically active conformation in a neutral salt/ethylene glycol mixture. BMP may be insolubilized by coprecipitation with calcium phosphate and resolubilized by chemical extraction with a neutral salt in the same solvent mixture. Upon concanavalin A-Sepharose chromatography, BMP is bound by hydrophobic interaction and carbohydrate recognition and is recovered by elution with either alpha-methyl mannoside or ethylene glycol solvent mixture.

Cartilage tissue differentiation from mesenchymal cells derived from mature muscle in tissue culture.

Under the influence of biochemical components of bone matrix gelatin (BMG), cartilage differentiates in tissue culture from the connective tissue cell outgrowths of mature muscle. Proliferation and differentiation begin within 24 hr with synthesis of hyaluronate, continue with high levels of synthesis of DNA and hyaluronidase, and culminate in production of large quantities of chondroitin sulfate. The addition of hyaluronic acid to the culture medium during the first 48 hr of culture depresses, whereas chondroitin sulfate enhances, subsequent production of cartilage.

Ultrastructure of chondrogenetic interactions between bone matrix gelatin and mesenchymal cells.

The ultrastructure of post fetal chondrogenesis was investigated in chondrogenetic implants and explants of bone matrix gelatin: (A) in direct contact with living cells; (B) separated from living cells by cellulose acetate membranes; (C) in serum or in a culture medium with no contact with living cells. Ruthenium red (RR) staining coarse granules free of interconnecting filaments aggregated in interstitial fluid and inside the surface of the bone matrix gelatin within 72 hours in systems with either direct or indirect contact with living connective tissue cells. No RR granules formed when control denatured or autolysed bone matrix gelatin were substituted for undenatured bone matrix gelatin.

Experimental myositis ossificans: cartilage and bone formation in muscle in response to a diffusible bone matrix-derived morphogen.

Bone matrix gelatin, prepared by chemical extraction of soluble noncollagenous proteins, was half digested with a chromatographically purified collagenase. The residue was placed on one side and autologous muscle on the other side of cellulose acetate membranes in diffusion chambers and tissue cultures. In this avascular system, the muscle septa connective tissue proliferated and differentiated into cartilage.

Müller's muscle-conjunctival resection ptosis procedure.

The Müller's muscle-conjunctival resection procedure is a relatively simple means of relieving ptosis in patients whose upper eyelids elevate to a normal level with phenylephrine instillation. The Müller's muscle-conjunctival resection procedure has advantages over the Fasanella operation because tarsus is preserved, and over the levator aponeurosis advancement and tuck operation because the results are more predictable.

Osteogenesis and chondrogenesis in transplants of Dunn and Ridgway osteosarcoma cell cultures.

Dunn osteosarcomas synthesize 2 times more alkaline phosphatase than do Ridgeway osteosarcomas, 3 times more than do HeLa cells, and 4 to 5 times more than do rat or mouse fibroblast cell cultures. Implants of killed freeze-dried Dunn cell cultures into the thigh muscles are resorbed and replaced by normal cartilage, bone, and bone marrow tissue, while implants of freeze-dried Ridgeway cells are resorbed and replaced by fibrous tissue only. Outgrowths of normal muscle septum connective tissue cells onto the stroma of Ridgeway tumors differentiate into fibrous tissue.

A rational approach to the surgical management of melanoma.

Our experience with 294 regional lymph node dissections in 250 patients are reviewed. The relationship between the Clark's level of invasion and the thickness of the primary is related to regional lymph node metastases. Patients with Clark's Level III melanoma had a 29% incidence of regional lymph node metastases, Clark's Level IV had a 42% incidence of regional lymph node metastases and Clark's Level V a 58% incidence of regional lymph node metastases.

Clinical observations in 101 consecutive patients with Duane's retraction syndrome.

We retrospectively studied 101 consecutive patients with Duane's retraction syndrome. Eighty-five percent were unilaterally affected and 15% were bilaterally affected. More patients had straight eyes in primary position than esotropia or exotropia.

The nature of immune complexes in human cancer sera.

Sera from 517 patients with various types of malignancies were assayed for immune complexes (ICs) by the Raji cell radioimmune assay. The incidence of immune complexes in these patients ranged from 16 to 52% as compared to 19% in normal controls. Increases in tumor mass and metastatic disease were associated with high levels of circulating ICs.

The effects of zinc deprivation on the host response to intramuscular bone matrix implants in the rat.

Sterile sequentially extracted insoluble bone matrix gelatin and freeze-dried allogeneic demineralized whole bone matrix was implanted into a lumbar muscle pouch of 12 Sprague-Dawley rats fed a diet containing 0.9 ppm zinc and into 6 rats fed a diet containing 50 ppm zinc. Histological sections and microradiography demonstrated that skeletal tissue induction occurred in all instances.

Transmembrane bone morphogenesis across multiple-walled diffusion chambers. New evidence for a diffusible bone morphogenetic property.

Bone generation and regeneration are associated with a bone morphogen that recruits mesenchymal cells for differentiation into bone. Experiments with particulate bone matrix gelatin implanted in multiple-walled diffusion chambers suggest that bone morphogen is a rapidly diffusible molecule, and consists of a noncollagenous bone morphogenetic protein (BMP). When particulate bone matrix gelatin is implanted inside of diffusion chambers constructed of two to five membranes, ranging from 300 to 750 cu micronm in total thickness, large deposits of bone develop on the outside.

An osteosarcoma cell and matrix retained morphogen for normal bone formation.

Histophysiology, ultrastructure, chemical analyses of transplants and implants of Dunn and Ridgway mouse osteosarcomas demonstrate that tumorigenesis is a manifestation of deranged morphogenesis in developing mesenchymal cell populations. The end product of development is defective, incompletely calcified, disorganized bone without any inclusions of bone marrow tissue. When Dunn osteosarcoma is freeze-dried and then implanted, the tumor is resorbed and replaced by deposits of normal cartilage, bone, and bone marrow.

Noncollagenous proteins of a rat dentin matrix possessing bone morphogenetic activity.

An insoluble preparation of rat dentin matrix was shown to possess bone morphogenetic protein (BMP) activity, i.e. the capacity to induce the formation of catilage and bone when implanted intramuscularly.

A bone matrix calcification-initiator noncollagenous protein.

When completely demineralized, the densely packed structure of bone matrix does not recalcify, neither in physiologic solutions in vitro nor in implants in vivo. Even when inorganic and organic calcification inhibitors (which normally are stored in bone matrix) are removed first by autolytic digestion in neutral buffers at 37C and then by sequential chemical extraction, implants of the EDTA insoluble residue will not recalcify after as long as 4 wk in a muscle pouch. However, if first demineralized in cold dilute HCl, second, extracted and autodigested in buffers solution at 37C, and then further extracted in EDTA and other solutions at 2C, a calcification initiator protein (Cp) is unmasked, and the residue will invariable recalcify.

In vitro evidence for increased cellular immunity to lung cancer antigen during Levamisole immunotherapy.

Levamisole, a potentiator of cell-mediated immunity, has been reported to increase survival in patients with resectable carcinoma of the lung. Cell-mediated immunity can be measured in vitro by the leukocyte migration inhibition test. We have previously reported that this test detects cell-mediated immunity to human lung tumor antigens.

Chondrogenesis in outgrowths of muscle tissue onto modified bone matrix in tissue culture.

Connective tissue outgrowths of muscle occur in vitro onto a substratum of bone matrix gelatin (BMG) chemically modified by various inorganic and organic cations or anions. Outgrowths onto BMG denatured by lyotropic concentrations of Ca2+ or complexed by only 1 mmole/1 of Cu2+ differentiate into fibrous tissue only. Onto BMG complexed with either positively or negatively charged organic molecules, the product was also fibrous tissue.

Collagenase-released non-collagenous proteins of cortical bone matrix.

Two distinct groups of non-collagenous components were isolated from rat cortical bone gelatin which had previously been digested with purified bacterial collagenase. One component was disulfide-bonded, strongly acidic, trypsin-labile glycoprotein aggregate with a molecular mass of more than 100,000 daltons. When reduced with beta-mercaptoethanol this protein disaggregated into subunits with a molecular mass of about 60,000 daltons.

Radioactive isotope labeled diffusible components of a bone morphogenetic substratum.

Bone matrix gelatin labeled with radioactive glucose, cysteine, proline, tyrosine or tryptophan was enclosed in one chamber while an adjoined second chamber was left empty to become filled with interstitial fluid. The paired diffusion chambers were implanted into the anterior abdominal wall of allogeneic rats for 14 days. Release of the matrix gelatin (BMG) from the loaded chamber was followed by a bone formation on the outer membrane of the unloaded chamber.

Reconstruction of the upper eyelid crease and fold.

For good cosmesis the upper eyelid creases and folds must be symmetric. The lid creases must be the same height above the upper lid margins. The skin below the crease must be smooth and firmly attached to the tarsus, or, in cases with a short tarsus, adherent to the short tarsus and levator muscle up to the crease.

Detection of human tumor-associated antigens by the leukocyte migration in agarose assay.

Twenty-three of 36 (64%) lung cancer patients, 19 of 36 (54%) melanoma patients and 18 of 27 (66%) sarcoma patients tested in the leukocyte migration in agarose assay against soluble extracts of histologically similar tumors showed significant inhibition of leukocyte migration. Reactivity to extracts of dissimilar tumors was low. Sera of only 1/13 (7%) lung cancer patients, 2/19 (10%) melanoma patients and 7/21 (33%) sarcoma patients were inhibited by extracts of histologically dissimilar tumors.