萬靈膏對骨折癒合療效的組織形態學研究

A Histomorphometric Study of Therapeutic Effect of Wan-Ling Paste on Fracture Healing

紐西蘭種雄性大白兔40雙（平均體重3公斤）將之分成四組，分別於術後一、二、三和四星期犧牲之。每隻動物右側脛骨(tibia)鑽一直徑3厘米的孔和右側排骨(fibula)鋸斷一長約3厘米的間隙，是對照組。左側經骨和腓骨做同樣處理，並以萬靈膏(wan-Ling Paste)外敷於手術部位的皮膚外表，是為萬靈膏組。動物於既定時間犧牲後，以骨組織形態學(bone histomorphometry)和顯微放射攝影學(microradiography)來觀察外表與鏡檢組織結構上的變化情形。本研究旨在探討萬靈膏對(1)骨折癒合過程中骨生成(bone formation)和骨吸收(bone resorption)的影響情形，和(2)骨組織的修復作用所扮演的角色。在脛骨鑽孔後一星期，從顯微放射攝影的觀察，對照組與萬靈膏組間沒有差異，但鏡檢發現在鑽孔處附近，萬靈膏組比照組具有較多的肉芽組織和較少的血凝塊。另外萬靈膏組在孔洞兩側皮質骨處，有破骨細胞和造骨細胞的出現，並且在封閉骨痂(sealing callus)處有織狀骨(woven bone)的形成，惟此等顯微結構的變化，卻未見於對照組中。此結果顯示，萬靈膏可加速骨折後癒合過程的進行，即縮短發炎時期(inflammatory stage)及加快進入修復時期(reparative stage)的過程。脛骨鑽孔後二星期，顯微放射攝影像的觀察，萬靈膏組具有較多的聯合骨痂(uniting callus)的形成。鏡檢發現，對照組在鑽孔兩側皮質骨處，有新的骨質生成，而萬靈膏組也見骨質生成，並有顯著增加的趨勢。另尚發現，萬靈膏組在鑽孔兩側皮質骨端及骨髓腔近封閉骨痂處，有多量骨質正在生成，比現象在對照並不明顯。此結果顯示，萬靈膏可加速骨折後癒合過程中修復時期的進行。鑽孔後三和四星期，顯微放射攝影像發現，萬靈膏組的骨修復過程比對照組快速，並且進入了重塑型時期(remodeling stage)。鏡檢發現，對照組繼續形成聯合骨痂，惟並未完全聯接兩側皮質骨。萬靈膏組聯合骨痂的形成已近完成，並幾全聯接兩側皮質骨。有關腓骨鋸斷後修復的情形，在術後一星期無論對照組或萬靈膏組均無變化。在術後二、三和四星期發現對照組和萬靈膏組均已在進行骨修復，惟萬靈膏組骨修復的進行速度和程度要比對照組來得快且好。綜上結果知，萬靈膏可刺激骨折處較多肉芽組織的生成以及各種骨痂的加速形成和骨生成的作用。簡言之，萬靈膏的使用，可促進骨折癒合(fracture healing)的效果。

Forty New-Zealand male rabbits, weighing 3.0kg, were randomly divided into 4 groups of 10 animals each. All rabbits received a 3mm in diameter drilled hole operation at the shaft 3cm underneath the superior fibular incisura of the proximal tibia by using 3mm in diameter round bone bur, and a 3mm long fracture operation at the shaft 2cm underneath the superior tibial incisura of the proximal fibula by using fissure bone bur on both hind limbs. The animal s were wrapt with placebo over the skin of operated area of right hind limb as the control group and with Wan-Ling Paste (WLP) over the left hind limb as the WLP group. Replacement of wrapt WLP twice a week for each animal. For experiment the animals were sacrificed at 1, 2, 3 or 4 weeks intervals after operation, respectively. The day and 13 days after hind-limb operation, all animals were received 25mg/kg of tetracycline HCI and 90mg/kg of xylenol orange, respectively. The tibia and fibula of each hind limb of the rabbit were removed whole and assayed microradiographically, contact microradiographically and histomorphometrically for fracture healing observations in callus and bone formation, bone resorption, bone cell kinetics and other elements in addition to gross appearance. No significant difference in the gross appearances of the control and WLP at l-week of treatment were observed. However, the histomorphometric structure of the WLP treated group showed more granulation tissue and less blood clot at the fracture site of the tibial cortex than the control group did. In addition, the osteoblasts and osteoclasts appeared at both fracture ends of the WLP treated group. These results indicated that WLP could cause a shortage of inflammatory stage of fracture healing. At 2-week WLP treated group, more uniting callus and bone formation were found between two fracture ends of the tibial cortex when compared to that of the control group indicating that WLP could a1so enhance the reparative stage of fracture healing under the condition. The uniting callus filled a1mostcompletely at two fracture ends of the tibial cortex in the 4-week WLP treated group. However, uniting callus was continuous forming but not connecting with the two fracture ends of the tibial cortex at the 4-week control group. The results indicated that processes of bone modeling and remodeling were proceeded better in WLP than that of the control group. At l-week, there were no significant changes observed microradiographically at the fracture site of fibula. However, at 2-,3-and 4-week of the WLP treated groups better reparative efforts were found when compared to that of the control group. Based on microradiographic, contact microradiographic and histomorphometric findings, it suggests that WLP not only increase the granulation formation, callus and bone formation but also promote the proceeding of bone modeling and remodeling. In conclusion, WLP could enhance the fracture healing.