PURPOSE: In the autologous fat injection, the centrifugation is useful for the refinement of harvested fat. As it can be an injury to the fat cell, we studied the fat cell viability with the change of centrifugation velocity and centrifugation time in order to get the limits of centrifugation velocity and centrifugation time. METHODS: We used the Colman System in 8 patients. We handled the control group with no centrifugation, group I with the centrifugation with 1500rpm for 1 minute, group II with 1500 rpm for 3 minutes, group III with 1500rpm for 5 minutes, group IV with 3000rpm for 1 minute, group V with 3000rpm for 3 minutes, group VI with 3000rpm for 5 minutes, group VII with 5000rpm for 1 minute, group VIII with 5000rpm for 3 minutes, group IX with 5000rpm for 5 minutes. We used the collagenase to separate the fat tissue. We had evaluated the fat cell viability by checking survival cell counts. RESULTS: There was no significance in group I, II, IV, V, but there was significant difference in group III, VI, VII, VIII, IX. CONCLUSION: The centrifugation with 3000rpm for 3 minutes is recommendable.
Adipocytes* ; Cell Count ; Centrifugation* ; Collagenases ; Humans

Keratocyne(R) is the complex of cystine and vitamine B6. Cystine is collagenase inhibitor and vitamine B6 is essential to the metabolism of cystine. We administered Keratocyne(R) orally combined with specific topical therapy to investigate its healing effect on various corneal diseases. The results were as follows: 1. The cases which improved after Keratocyne(R) administration were 15 patients out of 22 patients. 2. No untoward side effects were observed. We concluded that Keratocyne(R) was effective on certain corneal diseases which released collagenase.
Collagenases ; Corneal Diseases* ; Cystine ; Humans ; Metabolism ; Vitamins

New born rat heart cells were dissociated using trypsin and/or collagenase to elucidate the dissociation efficiency of these two enzymes. And the effect of dimethyl sulfoxide during and immediately after cell dissociation was also investigated to clarify the so-called protective activity of dimethyl sulfoxide on cell performance. The results can be summarized as follows. 1. Cold trypsin 18 hours pretreatment followed by warm collagenase treatment resulted best cell viability and cell yield. 2. Single warm trypsin treatment gave the poorest result. 3. Dimethyl sulfoxide did not seem to play any protective role during or immediately after rat heart cell dissociation. It had very damaging effect on rat heart cells.
Animals ; Cell Survival ; Collagenases* ; Dimethyl Sulfoxide* ; Heart* ; Rats* ; Trypsin*

The pathogenesis of scleroderma has not been completely delineated, but it is suggested that increased collagen expression in fibroblast from sclerotic skin lesions may be an important factor contributing to collagen accumulation. PUVA therapy has been reported to be effective in the treatment of localized scleroderma. Its mechanism of action appears to be the stimulation of collagenase production by dermal fibroblasts. We report a case of localized scleroderma improved with systemic PUVA therapy.
Collagen ; Collagenases ; Fibroblasts ; PUVA Therapy* ; Scleroderma, Localized* ; Skin

The author has observed the effects of collagenase on the relapse phenomenon and the histochemical changes during the relapse period. 50 rats were used. : 3rats as a normal group, 15rats as control groups, and 32rats as experimental groups. Rat's teeth were moved for 10days with helical spring applied, followed by injection of "collagenase in Hank's sol." to the experimental groups and the "Hank's sol." to the control group in the interdental gingiva on the 10th day, and the spring was removed on the 11th day. After injection, the experimental animals were sacrificed on the 11th, 13th, 15th, 17th, 20th, and 24th day and perpared histochemically for the Hematoylin-Eosin, Van-Gieson, and Methyl Green-pyronin staining. The results are as follows: 1. Group I (11th day): In the control group the supracrestal fibers were stretched and the metabolic rate was high. Experimental group showed that supracrestal fibers were resor, bed, disarrayed, and the metabolic rate was low. 2. Group II (13th day): In the control group, the supracrestal fibers began to vhange from the vertical direction to tooth-axis to the parallel. Experimental group showed that supracrestal fibers were completely resorbed. 3. Group IV (17th day): The control group showed almost normal structure. Form this group the metabolic rates were low. Experimental group showed the most destructive pattern. 4. Group VI (24th day): Experimental group showed almost normal structure. It follows that experimental groups were relapsed less than the control groups, and collagenase was effective in the prebention of relapse after rat's experimental tooth movement.
Animals ; Collagenases* ; Gingiva ; Rats ; Recurrence* ; Tooth Movement* ; Tooth*

The effect of orthodontic force on the collagenase and phosphatase activities of the adjacent alveolar bone was evaluated. Maxillary canines of male cats were treated orthodontically with closed coil spring so as to exert about 80g force. Sixteen cats were equally divided into one control group and seven experimental groups (12 hrs, 24 hrs, 36 hrs, 2 days, 3 days, 5 days and 7 days after orthodontic treatment). After sacrificing all animals on experimental intervals, collagenase, acid phosphatase (ACP) and alkaline phosphatase (ALP) activities were determined in the pressure and tension sides of alveolar bones. ACP activities increased in both the pressure and tension sides, but significantly increased in the pressure side continuously. ALP activities increased in the tension side at early stage (1-2 days after treatment), but changed small amount in the pressure side. Collagenase activities increased in the pressure side, especially at late stage (5-7 days after treatment). These results suggest that (1) orthodontic force increases the ACP, ALP and collagenase activities generally and (2) activities of ACP and collagenase increase in the pressure side, but that of ALP in the tension side and (3) activities of ACP and ALP increase at early stage, but that of collagenase at late stage after orthodontic treatment. Therefore it is shown that there are time differences in the formation and destruction of organic and inorganic components in the bone metabolism of alveolus with application of the orthodontic forces.
Acid Phosphatase ; Alkaline Phosphatase ; Animals ; Cats ; Collagenases ; Humans ; Male ; Metabolism

This study is to investigate the mechanism of inhibitory effect of imipramine on the calcium utilization in single cells isolated from canine detrusor. 2 mm thick smooth muscle chops were incubated in 0.12% collagenase solution at 36degreeC, and aerated with 95% O2/5% CO2, and then cell suspension was examined Acetylcholine (ACh) evoked a concentration-dependent contraction of the isolated detrusor cells in normal physiologic salt solution (PSS), and the ACh-induced contraction was significantly inhibited by imipramine. In Ca2+-free PSS, ACh-induced contraction was less than those in normal PSS and it was not affected by the pretreatment with imipramine. Ca2+-induced contraction in Ca2+-free PSS was supressed by imipramine, but addition of A 23187, a calcium ionophore, overcomed the inhibitory effect of imipramine. High potassium-depolarization (40 mM KCl) evoked cell contraction, which was inhibited by imipramine. Caffeine, a releasing agent of the stored Ca2+ from sarcoplasmic reticulum, evoked a contraction of the cells that was not blocked by the pretreatment with imipramine. These results suggest that imipramine inhibits the influx of calcium in the detrusor cells through both the receptor-operated- and voltage-gated-calcium channels, but does not affect the release of calcium from intracellular storage site.
Acetylcholine ; Caffeine ; Calcimycin ; Calcium* ; Collagenases ; Imipramine* ; Muscle, Smooth ; Sarcoplasmic Reticulum

Plasmin is an enzyme which plays an important role in the inflammatory process by activating vasoactive amine and lysis of fibrin. On the other hand, plasmin is also known to activate latent collagenase. Plasmin is an activated form of plasminogen which is stimulated by the plasminogen activator that exists in plasma and tissue. Gordon et al(1980) insisted that collagenase is important to the formation of corneal ulcer because it destroys the collagen which is the main component of the cornea. Berman et al(1980) reported that corneal tissue destruction by plasminogen activator-plasmin system can be a cause of corneal ulcer. We could obtain the following results by checking the plasmin activity in the tear of chronic corneal ulcer patients and necrotizing scleritis patients. 1. The plasmin activity in the tear was increased in all three chronic corneal ulcer patients in concentration of 1/16 sigma unit/ml to 1/8 sigma unit/ml. 2. There was no plasmin activity in the tear of the two necrotizing scleritis patients.
Collagen ; Collagenases ; Cornea ; Corneal Ulcer* ; Fibrin ; Fibrinolysin* ; Hand ; Humans ; Plasma ; Plasminogen ; Plasminogen Activators ; Scleritis ; Tears*

Whereas mesangial and epithelial cells from glomeruli are commonly grown in vitro, there has been major difficulties in developing homogenous cultures of human glomerular endothelial cells. This study defines the conditions for the reproducible isolation and growth of homogenous monolayers of human glomerular endothelial cells based on the method of Green DF et al published in 1992. Using the selective media and the sieving method, fibronectin was required as a surface matrix after adequate collagenase treatment, and endothelial cell growth factor and heparin was needed for the continuous growth of endothelial cells. The endothelial cell growth factor was isolated from the bovine hypothalamic extracts. Glomerular capillary endothelial cells exhibited a cobblestone morphology at confluence and stained homogenously with von Willebrand factor(factor VIII). The cytokeratin and the actin were not stained. This study might be helpful for in vitro study to know the biological characteristics of human glomerular endothelial cells under the predetermined condition.
Actins ; Cell Culture Techniques ; Collagenases ; Endothelial Cells* ; Epithelial Cells ; Fibronectins ; Heparin ; Humans* ; Keratins ; Population Characteristics

We isolated the strain MBL13 with high collagenase productivity from the soil of piled up animal bones. It was identified as Bacillus cereus. We purified and characterized Bacillus cereus collagenase (BCC). The molecular weight of BCC was 38.0 kDa and the optimum temperature and pH for the enzyme activity were 40 degrees C and 8.0 respectively. The enzyme was stable when the temperature was below 50 degrees C, but only retained 10% activity when kept at 60 degrees C for 1 h. The enzyme activity was stable between pH 7.0-8.5. Some metal ions such as Ca2+, Zn2+, Mg2+ enhanced the enzyme activity, and Cu2+ brought the obvious inhibition. In addition, EDTA and EGTA could inhibit the enzyme activity. We suggested that the purified enzyme was a member of the metalloproteases. Based on the experiment of substrate specificity, we found that the purified enzyme was bone collagenolytic protease, and had a much stronger capacity of hydrolysis for type I collagen than that for type II collagen and type III collagen. By BCC hydrolyzing bone collagen, we obtained polypeptides with different chain lengths. The comparative test indicated that the hydrolysis capacity of BBC was higher than that of standard type I collagenase. The results introduced a new strain and a novel collagenolytic protease for industrial enzyme.
Animals ; Bacillus cereus ; enzymology ; isolation & purification ; Bone and Bones ; microbiology ; Collagenases ; biosynthesis ; isolation & purification ; Hydrolysis ; Soil Microbiology