摘譯自 Prof. Norberto Roveri 諾勃特羅韋裡教授之 BioRepair Plus 科學研究報告 研究單位：義大利波隆那大學研究所LEBSC 1 1 1

BioRepair 修復牙齒琺瑯質牙膏 Prof. Norberto Roveri 諾勃特羅韋裡教授 I am pleased to present BioRepair® Plus to you. BioRepair® Plus, developed by Coswell Research Laboratories in collaboration with the LEBSC (Laboratory of Environmental and Biological Structural Chemistry) at the University of Bologna, Italy,is the first toothpaste based on bioactive microparticles which, thanks to their composition, are ableto penetrate into the microscopic defects on the enamel and dentine surfaces, performing an effective remineralising and reparative treatment. The LEBSC has been operating for thirty years and uses the most advanced techniques to study the chemical and biological aspects of mineralization processes of calcified tissues and, in particular,bone tissue.Enclosed is an in-depth scientific study which supports the effectiveness of BioRepair® and its MICROREPAIR® action. Used daily, BioRepair® will repair tooth enamel and will make your teeth more healthy – definitely something to smile about! Thank you for your time. Yours faithfully, Prof. Norberto Roveri Full Professor of General and Inorganic Chemestry “G. Ciamician” Department of Chemistry, University of Bologna 我很高興介紹BioRepair® Plus修復牙齒 琺瑯質牙膏給您 BioRepair® Plus是義大利COSWELL研 究實驗室與義大利波隆那大學環境和生 物結構化學實驗室合作研發的第一個牙 膏，其基礎上組成的生物活性微粒，能 滲透到牙齒琺瑯質的微觀缺陷和牙本質 表面，執行一個有效的再礦化和修復。 該環境和生物結構化學實驗室LEBSC已 運作了30年，採用最先進的技術研究化 學和生物方面的鈣化組織的礦化過程， 特別是對骨組織。以下是一種深入的科 學研究，其證實了BioRepair®的有效性 和MICROREPAIR®的作用。每日使 用，BioRepair®牙膏將修復牙齒琺瑯 質，並將使你的牙齒更健康 – 絕對是一 個值得微笑的東西！ 感謝你的時間。 此致 諾勃特羅韋裡教授 一般和無機化學教授 “灣恰米錢“化學系， 波隆那大學 2 2 2

Enamel 琺瑯質 Like vertebrate bone tissues teeth are a composite natural material, the main component of which is an inorganic carbonated Hydr oxyapatite phase, representing 65-70% of dentine by weight and over 90% of enamel by weight. Dental enamel forms the thin outer coating of our teeth and is considered to be the hardest and strongest of all biogenic materials (figure 1). 像脊椎動物的骨組織一樣牙齒是一 種天然的複合材料，其主要成分是 一種無機碳酸 - 羥基磷灰石層面， 佔 65-70％的牙本質重量和90％以 上琺瑯質的重量。 牙琺瑯質形成我們牙齒表面的薄薄 的塗層，被認為是最強硬的生物材 料（圖 1）。 3 3 3

圖 2b：掃描電鏡圖像：層棒狀羥基磷灰石微粒 ，形成琺瑯質（比例尺= 1微米）。 經許可轉載: Lowestan和S.韋納，“在生物礦化”， Tooth enamel is composed mainly of large, prismatic, highly crystalline hydroxyapatite microparticles , together with a very low protein component (figs. 2a and 2b). 牙琺瑯質主要由大的，稜形的高晶羥基 磷灰石微粒，連同一個非常低蛋白成份 組成（圖 2a和2b）。 Figure 2b: SEM image: layers of rod-like hydroxyapatite microparticles that form enamel (Scale bar=1μm). Reproduced with permission from H. Lowestan and S. Weiner, "On Biomineralization", Oxford University Press 1989. 圖 2b：掃描電鏡圖像：層棒狀羥基磷灰石微粒 ，形成琺瑯質（比例尺= 1微米）。 經許可轉載: Lowestan和S.韋納，“在生物礦化”， 牛津大學出版社1989年。 Figure 2a: Enamel structure viewed with Scanning electron microscopy (SEM) (Scale bar=5μm). Reproduced with permission from H. Lowestan and S. Weiner, "On Biomineralization", Oxford University Press 1989. 圖 2a：琺瑯質結構效果掃描 電子顯微鏡（SEM）（比例尺=5μm的）。 經許可轉載: Lowestan和S.韋納， “在生物礦化”，牛津大學 1989年。 4 4 4

圖 3：，在使用掃描電子顯微鏡顯現的牙本質結構的效果（SEM）。在牙本質小管，可清楚地看到，它們經過成齒質細胞處理和管內牽連的流體所佔據。 In adults, enamel does not contain any cells and is therefore not capable of self regeneration. Any damage is irreversible as there is no biological process capable of repairing damaged enamel. In a similar way, dentine(figure 3) exposed to the oral environment cannot be regenerated, as new dentine is deposited on the interior surfaces of the crown close to the pulp, and not on the outside. For this reason, any reparative action must be provided by materials or substances that are extraneous to dental tissue metabolism. These substances are either synthetic or are precipitated from saliva 在成人，琺瑯質不包含任何細胞，因此 不能夠自我再生。任何損害不可逆轉 的，因為目前沒有生物過程能夠修復受 損的琺瑯質。 以類似的方式，牙本質（圖 3）暴露在 口腔環境，不能再生的，新的牙本質沉 積在接近牙骨髓的齒冠的內部表層上， 而不是在外面。為此，任何修復行動的 材料或物質其是與牙齒組織的代謝無 關。這些物質要麼是合成的或是從唾液 的沉澱中獲得。 Figure 3: Dentine structure viewed using Scanning electron microscopy (SEM). The dentinal tubules can be seen clearly, occupied by the odontoblast processes and intratubular fluid implicated. 圖 3：，在使用掃描電子顯微鏡顯現的牙本質結構的效果（SEM）。在牙本質小管，可清楚地看到，它們經過成齒質細胞處理和管內牽連的流體所佔據。 5 5 5

MICROREPAIR® BioRepair® Microparticles Ca(10-x)Znx(PO4)(6-y)(CO3)y (OH)2 (Y=4-8 % CO32- substituted at PO43-) (X=1 % Zn2+ substituted at Ca2+) BioRepair ®微粒 鈣（10 - x）的Znx（PO4）（6 - Y）的（碳酸 攝）（OH）2的（是= 4-8％碳酸 2 – 取代在PO4 3 - ）（x = 1的％Zn2 +的取代的鈣） Figure 4: Chemical formula for dentine and enamel remineralising hydroxyapatite microparticles (Microrepair®). Calcium phosphate and carbonate are present in very similar relationships to those occurring naturally. It is the zinc, with its antisepticproperties, that is responsible for the plaque-preventing action. 圖 4：牙本質和琺瑯質再礦化形成 羥基磷灰石微粒（Microrepair ®） 的化學公式。磷酸鈣和碳酸鈣與那 些自然產生的物質存在著非常類似 的關係。鋅的抗菌特性主要是負責 預防牙菌斑的侵入行動。 MICROREPAIR® The hydroxyapatite microparticles (MICROREPAIR®) present in BioRepair® Plus are completely identical to the mineral that forms dentine and enamel. It is precisely from this similarity that the synthetic micro particles derive their capacity to reconstruct dentine and Enamel. MICROREPAIR ® 存在於BioRepair® Plus的羥基磷灰 石微粒（MICROREPAIR ®）與構 成牙本質和琺瑯質的礦物是完全相 同的。正是從這個相似，合成的微 粒獲得重建牙本質和琺瑯質的能 力。 . 6 6 6

用於 BioRepair ® Plus 的物質是技術上的創 新，它以微粒的形式加 強了它的化學反應。微 粒以釋出其組成的鈣、 磷來履行牙本質的礦化 行動。微粒以他們能附 著在琺瑯質的自然組織 的能力來填補細微裂縫 的琺瑯質。 The substance used in BioRepair® Plus is technologically innovative, in that being in the form of microparticles it has enhanced chemical reactivity. Microparticles carry out their dentine remineralising action by releasing their constituent calcium and phosphorous in situ. In the case of the enamel, the microparticles action takes place via their ability to bond to natural tissues, thus filling microgaps in the enamel.。 7 7 7

Microscopic Microrepair® micro-particles aggregates viewed using TEM 圖 5和6： 微觀 Microrepair ® 微粒聚集效果，請使用透射電鏡 （透射電子顯微鏡） （比例尺= 100微米） Figures 5 and 6: Microscopic Microrepair® micro-particles aggregates viewed using TEM (Transmission electron microscopy) (Scale bar=100 μm) 8 8 8

(Transmission electron microscopy) (Scale bar=100 μm) Figures 7 and 8: Microscopic Microrepair® microparticles aggregates viewed using TEM (Transmission electron microscopy) (Scale bar=50 and 20 μm respectively). （透射電子顯微鏡）（比例尺= 100微米） 圖 7和8：微觀 Microrepair ®微粒聚集效果請使用透射電子顯微鏡 (透射電子顯微鏡) （比例尺= 50，分別為 20微米）。 9 9 9

Introduction介紹 Action mechanism The enhanced reactivity of MICROREPAIR® microparticles is due primarily to the bio-mimetic action of these microparticles, which have the distinguishing feature of possessing a very similar chemical composition to that of enamel and dentine. The X-ray diffraction spectrum of MICROREPAIR® microparticles (figure 9) shows how the degree of crystallinity of the microparticles is half way between that of enamel and dentine. 作用機制 增強反應的MICROREPAIR ®微粒 主要是由於對生物模仿動作，這些具有顯著特徵的微粒與組成琺瑯質和牙本質的化學成分相似。X射線衍射譜MICROREPAIR ®微粒（圖 9）顯示微粒的結晶度是介於琺瑯質和牙本質的中間。 Figure 9: X-ray diffraction diagrams of enamel (red line), MICROREPAIR® microparticles (blue line) and dentine (green line). 圖 9：琺瑯質的X -射線衍射圖（紅線）， MICROREPAIR ®微粒（藍線）和牙本質（綠線）。 10 10 10

使硬組織再礦化的能力會消除 對牙齒硬組織造成影響的障礙 物如： The ability to remineralise hard tissues has an impact on several disorders that affect the hard tissue of teeth: 1. Prevention of cavities by remineralisation of original lesions; 2. Desensitising effect on dentine by covering up dentinal tubules; 3. Tartar and plaque prevention thanks to antibacterial action of Zn2+, with recognized antiseptic properties 使硬組織再礦化的能力會消除 對牙齒硬組織造成影響的障礙 物如： 1 原組織的再礦化能預防蛀牙; 2 掩蓋牙本質小管，使牙本質 有抗敏作用; 3 具有公認的防腐性能的鋅， 其本身的抗菌作用能預防牙 結石和牙菌斑。 11 11 11

Figure 10: the SEM images show the surface of the dentine before treatment (a) and after treatment with MICROREPAIR®: 1 minute (b), 10 minutes © and one hour (d). They clearly show the progressive growth of the apatite microparticles until they completely hide the dentine channels 圖 10：在SEM圖像顯示未經過處理的牙本質表面（a）和用 MICROREPAIR ®處理後的1分鐘（b），10分鐘（c） 及1個小時（d）。他們清楚地顯示了磷灰石微粒的增長過程，直到他們完全掩蓋牙本質渠道 Figure 10a Figure 10 b Figure 10 c Figure 10 d 12 12 12

一個類似的機制可以看出琺瑯 質的表面在有微小的缺陷或簡 單的不規則表面，BioRepair ®Plus微粒如何來開始他們的 再結晶過程。 Figures 10 (a, b, c and d) show the progressive action of MICROREPAIR® microparticles, as they gradually bond securely to the surface of the dentine, blocking the tubules and thus performing their effective, long-lasting desensitising effect on the dentine. The phenomenon can be seen at work a few minutes after the BioRepair® Plus microparticles have been applied, which means that the microparticles remineralising and desensitising action begins after only a few applications of BioRepair® Plus toothpaste. A similar mechanism can be seen on the surface of the enamel where there are microscopic defects or simple surface irregularities, as the BioRepair® Plus microparticles are laid down and begin their recrystallisation process. 圖 10（的A，B，C和D）顯 示MICROREPAIR ®微粒的漸 進行動 ，因為他們逐漸安全 地附著在牙本質表面，阻塞微 小管，從而履行其有效且持久 的讓牙本質產生抗敏的作用。 在使用BioRepair ®Plus微粒 幾分鐘後就可看出上述的現 象，這意味著微粒再礦化和抗 敏行動開始後，只需使用適量 的BioRepair ®Plus牙膏。 一個類似的機制可以看出琺瑯 質的表面在有微小的缺陷或簡 單的不規則表面，BioRepair ®Plus微粒如何來開始他們的 再結晶過程。 13 13 13

鋅能釋放一個最佳濃度， 因此，能履行其在口腔內 的防腐劑行動。 The Zinc release mechanism is extremely innovative, as it takes place via slow dissolution of the apatite, which then releases its various components locally (Zn2+, Ca2+, CO3 2-, PO4 3-). The Zinc, in its ionic form as a bivalent action, then performs its antiseptic function inside the mouth. The Zinc is released at an optimum concentration, therefore carrying out its antiseptic action in the mouth. 鋅釋放機制是極具創新性 的，因為它是經由磷灰石 緩慢的溶解而發生的，然 後釋放其原本不同的成份 （鋅，鈣，碳酸2 - ，PO4 3 - ）。鋅離子以其雙向行 動的形式來執行它在口腔 內的防腐功能。 鋅能釋放一個最佳濃度， 因此，能履行其在口腔內 的防腐劑行動。 14 14 14

由於BioRepair ® Plus的微粒 具有吸收的特性，每日使用 Microrepair ®能有效地對付 口腔異味的問題。 4. Finally, apatite has other secondary effects, including the absorption of sulphate compounds such as H2S, which is responsible for halitosis. Daily use of BioRepair® Plus is therefore also indicated for control of this condition: thanks to the absorbent properties of BioRepair® Plus microparticles , daily use of Microrepair® is effective in fighting halitosis problems. 4 最後，磷灰石還有其他影 響，包括吸收硫酸鹽化合 物，如硫化氫，這是口臭 之原因。因此每天使用 BioRepair ® Plus就還能控 制這種情況： 由於BioRepair ® Plus的微粒 具有吸收的特性，每日使用 Microrepair ®能有效地對付 口腔異味的問題。 15 15 15

Fig.7 X-ray diffraction pattern of natural carbonate-hydroxyapatite from deproteinated dentine (a), synthetic CHA about 20 nm sized plate shaped microparticles (b), synthetic CHA about 100 nm sized plate-acicular shaped microparticles (c), and natural carbonate-hydroxyapatite from enamel (d). 圖 7脫蛋白牙本質的自然碳酸羥基磷灰石的X射線衍射模式（a），合成的CHA約 20奈米大小的微粒板形（b），合成的CHA約 100奈米大小的板形微粒針狀（c），和琺瑯質的自然碳酸羥基磷灰石（d）。 16 16 16

Current and future developments 現在與未來的發展 From some decades the teeth health care has been committed to the fluoride effect on hydroxyapatite. Fluoride interacts with hydroxyapatite promoting the conversion into fluorapatite, hich is less soluble and more mechanically resistant, but also more brittle than hydroxyapatite. The action of fluoride on the surface enamel apatite appears ideal to prevent demineralization due to acid food and drinks and to contrast plaque damage. A lot of scientific works support the fluoride positive action for teeth health care and consequently a lot of fluoridated oral care products have been patented till nowadays and in some countries drinking water has been enriched with fluoride. 過去幾十年來牙齒保健一直訴求 於氟對羥基磷灰石(即琺琅質)的影 響。 交互影響轉化後的氟羥基磷灰 石，是較不易溶解、更多的機械 抗阻性的、也比羥基磷灰石更易 碎。 使用氟對琺瑯質的表層毫無疑問 地似乎只是用來防止因酸食品和 飲料而脫礦化，和對抗牙菌斑的 損傷。目前還有很多科學工程支 持氟使用在牙齒保健，因此，直 到現在，仍然還有不少的含氟口 腔護理產品獲得專利而且在一些 國家飲用水也加入氟之成份。 17

歐洲食品安全局（EFSA）的科學小 組已建議有關氟攝入的風險並而對口 腔護理產品和牙膏設定最高的氟含量 水平為1500毫克/公斤。 Fluoride is not essential for human growth and its content in the body is not under physiological control. Adsorbed fluoride is rapidly distributed in the body by circulation and is retained only in calcified tissues. In the last decades dangerous effects in human health have been ascribed to the daily fluoride amount ingested and many studies have put in evidence its high risk, especially for children, to get fluorosis and bone diseasesin old people. European Food Safety Authority (EFSA) Scientific Panel has advised about the risk of fluoride intake from ingested oral care products and toothpastes can be fluoridated with a maximum level of1500 mg/Kg. In spite of this knowledge, fluoride continues to be widely used in orthodontia and dental applications to prevent enamel surface demineralization and represents the main component in toothpastes claiming remineralization effects. If nowadays fluoride represents the unique agent to contrast caries, plaques and demineralization we may probably ascribe this situation to the expensive cost of odontoiatric products containing hydroxyapatite. In spite of this, hydroxyapatite is commonly considered the main synthetic biomaterial as bone filler and substitute and only few daily use products for teeth health care have been patented. 氟化物對人的成長並無重要性，而它 的含量在體內是無法用生理來控制 的。吸附氟迅速分佈於體內的循環， 並且只保留在鈣化組織。在過去幾十 年危害人類健康的因素，也被歸咎到 與每日攝取的氟量有關。許多研究證 據證明氟的高風險性， 尤其是兒童會 得到氟而中毒，而老人則會有骨骼疾 病。 歐洲食品安全局（EFSA）的科學小 組已建議有關氟攝入的風險並而對口 腔護理產品和牙膏設定最高的氟含量 水平為1500毫克/公斤。 儘管已知這些知識，氟化物卻仍繼續 廣泛應用於牙齒矯正和防止牙釉質表 面脫酸蝕，而且被認為無以替代。這 種情況，我們也只能把它歸咎於羥基 磷灰石的產品成本昂貴，大家只好退 而求其次，選用有極大缺點的氟化 物，做為護齒的重要成份。 儘管如此，羥基磷灰石還是普遍的被 認為，對骨的填充物及代替品，它才 是最佳的合成生物材料。 18 18 18

過去都經常性地注入一個低量 的、較便宜的氟化物，用來改善 羥基磷灰石表面反應性。 Too expensive is the hydroxyapatite cost to be consistently utilised as an active agent in toothpastes or mouth washes. In fact, almost all the patents previously reported and highlighting the ability of hydroxyapatite to remineralize enamel and dentine, propose different compositions containing calcium and phosphate ions which can produce hydroxyapatite on teeth if mouth pH induces this crystallization and frequently are obliged to insert a low amount of fluoride in order to promote the apatite crystallisation. Small amounts of industrial hydroxyapatite are associated to other cheaper components and submitted to milling to improve hydroxyapatite surface area and reactivity. Only recently the development of nanotechnologies has opened new opportunities in obtaining cheap hydroxyapatite micro-nano particles by the “bottom up” methods. These hydroxyapatites are surface nanostructured and have higher surface area and consequently higherreactivity allowing them to bing to enamel and dentine apatite producing on enamel a biomimetic coating,contrasting plaque formation and sealing dentine tubules annulling hypersensibility. 太昂貴的成本，造成羥基磷灰石 一直無法被積極有效的利用在牙 膏和漱口水中。事實上，幾乎所 有以前的專利報告都揭示了羥基 磷灰石對琺瑯質和牙本質再礦化 的能力， 並提出了含有鈣和磷酸 離子不同的成份， 可在牙齒表面 產生羥基磷灰石， 並藉由口中的 PH質能誘導結晶。 過去都經常性地注入一個低量 的、較便宜的氟化物，用來改善 羥基磷灰石表面反應性。 直到最近，奈米技術的發展開設 了新的機遇，介由“由下往上”的 方法來獲得較經濟的羥基磷灰石 微奈米粒子 。 這些羥基磷灰石具有奈米結構化 的表面，且具有較高的表面面積 反應性，使它們能夠固定在琺瑯 質和牙本質的磷灰石上，並在琺 瑯質上產生一仿生表層，來對抗 牙菌斑的形成，及封閉牙本質的 小管，來對抗牙齒之過敏。 19