산골이 가골 및 골형성에 미치는 영향에 대한 실험적 연구

Abstract

[한글]

골절치유 기간을 단축시키고 골유합을 촉진시키기 위하여 오래전부터 많은 학자들에 의하여 가골형성 및 골유합에 관한 연구가 계속되고 있다. 그러나 현재로서는 골절치유를 촉진시킬 수 있는 확실한 방법은 없다고 하여도 과언은 아니다.

Key 및 Odell (1955)은 과량의 광물질 투여가 실험동물의 골절치유 과정에 별 영향이 없다고 보고하였으며 Kowalewski (1958)와 Hulth 및 Olerud (1964)등은 콜티손이 골절유합을 지연시킴을 증명한 바 있다. Koskinen (1963)은 성장홀몬과 갑상선 홀몬이 골절치유를 촉진시킨다고 하였으며 특히 지연유합이나 불유합에서 효과가 좋았다고 보고한 바 있다. 최근 Ziegler 및 Delling (1972)은 동물실험에서 칼시토닌(calcitonin)이 골재생 및 치유과정을 촉진함을 관찰 보고하였다.

Friedenberg등 (1971)은 전류자극으로서 골절치유를 촉진시키는 실험 보고를 발표한 바 있으며 Lavine등 (1971, 1972)은 전류자극으로서 선천성 경골 가관절증(pseudoarthrosis of tibia)을 성공적으로 치유시켰다고 임상보고를 한바도 있다.

우리나라에서는 언제부터 였는지는 알 수 없으나 산골이 골질환 및 골절치유에 효과가 있는 약제로서 민간 한방에서 사용되고 있다. 저자는 산골이 골절치유에 미치는 영향의 유무와 만일 영향이 있다면 그 양상의 일단을 추구코져 본 실험을 시도하였다.

본 실험을 시행함에 있어 1. 산골 성분에 대한 재분석. 2. 산골의 위장내 흡수 여부. 3. 장내 Ca**45(Ca**++)의 이동에 미치는 영향. 4. 흡수된 산골의 각조직에의 분포상태. 5. 산골이 골절조직의 치유과정에 미치는 영향 등으로 구분하여 실험하였고 특히 산골의

골절된 조직에서의 치유과정에 미치는 영향을 보기 위하여 1). 산골이 유기기질 형성기에서 sulfate 고착에 미치는 영향. 2). 이 과정에서의 조직학적 검사. 3). 석회화(calcification)과정에 밀접한 관계가 있는 alkaline phosphatase 활성의 조직화학적 검사를 하여 비교관찰하였다.

실험방법으로 산골의 장내흡수 실험을 위하여 12마리 가토의 공장분절(jejunum segment)과 5마리의 쥐의 위를 사용하여 incubation bath에 장치한 다음 실험하였고 흡수된 산골의 각조직에의 분포 상태를 보기 위하여 5마리의 쥐에 방사성화된 산골의 일정량을 도관

급식(tube feeding)을 시킨다음 각 장기를 채취하여 이를 완전히 태운후 산골의 방사능을 측정하였다.

산골이 골절조직의 치유과정에 미치는 영향을 보기 위하여 한국산 성숙 웅성 가토 106마리의 양측 비골을 이용하였으며 실험동물은 30mg/kg의 nembutal 정주(靜注) 마취하에 무균적으로 동일한 골절을 만들 목적으로 치과용 회전 disc을 사용하여 비골 골두 하방 1.0cm 부위에 횡적골적을 일으킨후 마취에서 깨어나는 대로 부목 고정없이 보행토록 하였다.

실험동물을 2군으로 구분하여 그중 1군에서 술후 1일부터 산골 1gm을 교갑에 넣어 계속 투여하여 산골투여군으로 하였으며 비투여군을 대조군으로 하였다. 각군은 다시 4일과 1,2,3,4,5,7 및 12주로 구분하여 공기주입으로 희생시킨 다음 육안적 소견, X-선 검사, 조직학적 검사 및 alkaline phosphatase 활성의 조직화학적 검사를 하여 비교 관찰하였다.

골절치유 과정중 유기기질 형성기에 있어서 sulfate 고착을 보기 위하여 실험동물에게 S**35 (Na^^2S**35O^^4) 동위원소를 매일 20μci씩 복강내 주사하고 6일째 동물을 희생시킨 다음 골절골에 고착되어 있는 S**35O^^4-- 방사능을 scintillation spectromenter로

측정하여 비교 관찰하였다.

실험결과

1. 산골은 토끼소장에서 Ca**45(Ca**++)의 이동에 영향을 주지 않았다.

2. 산골은 적출된 쥐의 위에서 흡수되었다.

3. 흡수된 산골은 타조직보다 간조직에 의의있게 많이 분포되었다.

4. 산골은 골절치유 과정에서 유기기질 형성시기에 S**35O^^4-- 고착에 영향을 주지 않았다.

5. 골절치유 과정중 육안 및 X-선 검사소견상 산골투여군에서 시기적으로 골유합이 조기에 나타났다.

6. 조직학적 검사소견에서 산골투여군은 조기에 가골형성이 대조군보다 더 빨리 또한 왕성하게 진행되었다.

7. Alkaline phosphatase 활성은 산골투여군에서 대조군에 비하여 특히 조기에 강하고 광범위하게 나타났다.

이상의 결과로 미루어 산골은 위에서 흡수되며 골절골의 치유과정 중 특히 조기에 있어서 석회화 과정에 관여하여 골절치유를 촉진하는 것으로 사려된다.

An Experimental Study on the Effect of "Sangol" upon Callus and Bone Formation

Hun Sik Choi, M.D.

Department of Medical Science, The Graduate School, Yonsei University

(Directed by Prof. In Hee Chung)

There are numerou sexperimental studies for stimulation of fracture healing by

means of administration of minerals, hormones, vitamins, and electric energy. At

present, no direct method of controlling the healing of fractured bone exists.

Key and Odell(1955) found that excess minerals did not accelerate the healing of

experimental fractures in rats. The effect of cortisone on fracture healing was

studied by Kowalewski(1958) who found with hemerus fractures in rats that the

uptake of S**35 was considerably less than in normal fracture. Hulth and Olerud

(1964) found that the most obvious difference between the normal and the

cortisone-treated fractures was the delayed reconstruction of fracture hematoma in

the latter. Ziegler and Delling (1972) attempted to identify a positive calcitonin

effect in holes bored in the cortical bone of 40 female Sprague-Dawley rats. They

found that the calcitonin effect of acceleration of the healing process was

demonstrable only within the first 3-4 weeks. Conchran (1972) said that physiologic

electric currents existed in living bone and apparantly acted in bone remodelling

as well as in fracture healing. Lavine, et al. (1972) reported a case of human

congenital pseudoarthrosis of the tibia, which had been treated successfully with

electric current.

"Sangol" means "Bone of Mountain" in Chinese letters. It is a mineral composed

mainly of Fe and S, and its minor constituents are Si, Al, Mg, Pb, Mn, Cu, Na, Ti,

and Ca.

In korea, it is being used in bone diseases, especially in fractures of bones by

Korean herb doctors, without scientific proof. Therefore, the purpose of this

experimental study is to find its effect upon the healing of fractures.

The experimental plans were as follows: 1). re-analysis of the chemical

composition of the Sangol. 2). absorption of the Sangol. 3). its effect on Ca**45

(Ca**++) transport across the rabbit's jejunum. 4). distribution of the Sangol to

each organ. 5). effect of the Sangol upon the healing of experimental fractures in

rabbits; a). effect of the Sangol on S**35O^^4-- fixation in fractured bone tissue.

b). roentgenographic and macroscopic observation. c). histological examination (H-E

stain). d). histochemical study for the activity of alkaline phosphatase.

One hundred and eighteen albino rabbits and ten albino rats were used in this

experiment. Twelve of the rabbits and ten rats were used to study the absorption of

Sangol. The remaining 106 rabbits were operated in sterile conditions under

intravenous nembutal anesthesia (30mg per kg of body weight). Both fibulae were

exposed through antero-lateral skin incisions, and clean transverse shaft fractures

were made 1.0cm below the heads of the fibulae using a fine rotating dental disc.

The operated rabbits were divided into two groups; the first group (experimental

group, 53 rabbits) was medicated with 1gm of Sangol mixed with normal laboratory

diet, and the second group (control group, 53 rabbits) was fed only with regular

laboratory diet.

The animals of each group were sacrificed by air embolization in four days, and

1,2,3,4,5,7 and 12 weeks after fracture in 8 subgroups.

The results of the experiment are as follows;

1. Sangol did not influence the Ca**45(Ca**++) transport across the rabbit's

jejunum.

2. Sangol was absorbed in rat gastric mucosa.

3. Sangol was distributed significantly in the liver.

4. Sangol did not influence the S**35O^^4**-- fixation in fractured bone.

5. On macroscopical and roentgenographical examination, callus formation and bony

union of the medicated group occurred earlier than in the control group.

6. On histological examination, the callus formation and bony union of the

medicated rabbits occurred much more rapidly and vigorously, especially in the

early stage of fracture healing, than in those of the control group.

7. On histochemical examination for the activity of alkaline phosphatase,

particularly in the early process of fracture healing, a stronger and wider

reaction was observed in the slide preparations of the medicated group than in the

corresponding ones of the control group.

These results suggest that Sangol can be absorbed in the stomach and may promote

and accelerate the calcification process of fracture healing with increasing

activity of alkaline phosphatase.

[영문]

There are numerou sexperimental studies for stimulation of fracture healing by means of administration of minerals, hormones, vitamins, and electric energy. At present, no direct method of controlling the healing of fractured bone exists.

Key and Odell(1955) found that excess minerals did not accelerate the healing of experimental fractures in rats. The effect of cortisone on fracture healing was studied by Kowalewski(1958) who found with hemerus fractures in rats that the uptake of S**35 was considerably less than in normal fracture. Hulth and Olerud (1964) found that the most obvious difference between the normal and the cortisone-treated fractures was the delayed reconstruction of fracture hematoma in the latter. Ziegler and Delling (1972) attempted to identify a positive calcitonin effect in holes bored in the cortical bone of 40 female Sprague-Dawley rats. They found that the calcitonin effect of acceleration of the healing process was demonstrable only within the first 3-4 weeks. Conchran (1972) said that physiologic electric currents existed in living bone and apparantly acted in bone remodelling as well as in fracture healing. Lavine, et al. (1972) reported a case of human

congenital pseudoarthrosis of the tibia, which had been treated successfully with electric current.

"Sangol" means "Bone of Mountain" in Chinese letters. It is a mineral composed mainly of Fe and S, and its minor constituents are Si, Al, Mg, Pb, Mn, Cu, Na, Ti, and Ca.

In korea, it is being used in bone diseases, especially in fractures of bones by Korean herb doctors, without scientific proof. Therefore, the purpose of this experimental study is to find its effect upon the healing of fractures.

The experimental plans were as follows: 1). re-analysis of the chemical composition of the Sangol. 2). absorption of the Sangol. 3). its effect on Ca**45 (Ca**++) transport across the rabbit's jejunum. 4). distribution of the Sangol to each organ. 5). effect of the Sangol upon the healing of experimental fractures in

rabbits; a). effect of the Sangol on S**35O^^4-- fixation in fractured bone tissue.

b). roentgenographic and macroscopic observation. c). histological examination (H-E stain). d). histochemical study for the activity of alkaline phosphatase.

One hundred and eighteen albino rabbits and ten albino rats were used in this experiment. Twelve of the rabbits and ten rats were used to study the absorption of Sangol. The remaining 106 rabbits were operated in sterile conditions under intravenous nembutal anesthesia (30mg per kg of body weight). Both fibulae were exposed through antero-lateral skin incisions, and clean transverse shaft fractures were made 1.0cm below the heads of the fibulae using a fine rotating dental disc.

The operated rabbits were divided into two groups; the first group (experimental group, 53 rabbits) was medicated with 1gm of Sangol mixed with normal laboratory diet, and the second group (control group, 53 rabbits) was fed only with regular laboratory diet.

The animals of each group were sacrificed by air embolization in four days, and 1,2,3,4,5,7 and 12 weeks after fracture in 8 subgroups.

The results of the experiment are as follows;

1. Sangol did not influence the Ca**45(Ca**++) transport across the rabbit's jejunum.

2. Sangol was absorbed in rat gastric mucosa.

3. Sangol was distributed significantly in the liver.

4. Sangol did not influence the S**35O^^4**-- fixation in fractured bone.

5. On macroscopical and roentgenographical examination, callus formation and bony union of the medicated group occurred earlier than in the control group.

6. On histological examination, the callus formation and bony union of the medicated rabbits occurred much more rapidly and vigorously, especially in the early stage of fracture healing, than in those of the control group.

7. On histochemical examination for the activity of alkaline phosphatase, particularly in the early process of fracture healing, a stronger and wider reaction was observed in the slide preparations of the medicated group than in the corresponding ones of the control group.

These results suggest that Sangol can be absorbed in the stomach and may promote and accelerate the calcification process of fracture healing with increasing activity of alkaline phosphatase.