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Table of Contents
ORIGINAL ARTICLE
Year : 2021  |  Volume : 70  |  Issue : 2  |  Page : 93-96

Comparison of maceration techniques for retrieval of bones


Department of Anatomy, Dayanand Medical College and Hospital, Ludhiana, Punjab, India

Date of Submission12-Aug-2020
Date of Acceptance28-Apr-2021
Date of Web Publication30-Jun-2021

Correspondence Address:
Mrs. Anshu Soni
Asst.Prof, Department of Anatomy, Dayanand Medical College and Hospital Ludhiana, Punjab
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/JASI.JASI_47_20

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  Abstract 


Introduction: Learning osteology is an essential part of anatomy curriculum. However, recently, there has been a scarcity in the availability of bones for teaching in India. The present study was conducted to compare four different maceration techniques and find the most suitable method to procure bones from an embalmed cadaver. The present study was conducted on dissected human cadavers made available from the department of anatomy. Material and Methods: For the first three methods, after dissection, the cadavers were buried in the burial section of the department for 2 years. The bones were washed after excavation. In the first method, detergent was used; in the second, quick lime was used; and in the third, hydrogen peroxide was used. In the fourth method, parts were not buried. The dissected parts were defleshed manually. The bones were freed from the joint. A paste was made using baking soda and 30% hydrogen peroxide. This was applied and bones were kept covered for 24 h. All four methods were compared. Results: Based on time required to macerate, ease of soft-tissue removal and complete procurement with minimal damage, the method using 30% H2O2 was the best. For smaller specimens, the new method of applying a paste of baking soda and H2O2 was very effective. The time consumed was very less. Discussion and Conclusion: Maceration using 30% H2O2 gave the best result. New method tried is time-saving and useful for small specimens. Soft tissue was removed easily with no/minimal damage to the bone.

Keywords: Baking soda, bones, embalmed, hydrogen peroxide, maceration, quick lime


How to cite this article:
Soni A, Kumar A, Sharma A, Vohra H. Comparison of maceration techniques for retrieval of bones. J Anat Soc India 2021;70:93-6

How to cite this URL:
Soni A, Kumar A, Sharma A, Vohra H. Comparison of maceration techniques for retrieval of bones. J Anat Soc India [serial online] 2021 [cited 2021 Jul 31];70:93-6. Available from: https://www.jasi.org.in/text.asp?2021/70/2/93/320260




  Introduction Top


Learning osteology is an important part of the anatomy curriculum. While modes such as computer applications, textbooks, and plastic models help students to learn how the human body works, there is virtually no substitute for the use of authentic human tissue. Human bones are unsurpassed in the ability to provide three-dimensional instruction in osteology as well as understanding the soft-tissue insertion and the course of neurovascular structures.[1] The normal variations in the human anatomy cannot be adequately reproduced in software, illustrations, or models. Even photographs, while useful as documentation, cannot replace the tactile appreciation of bone. While nonhuman remains are readily available, without access to human skeletal resources, a comparative study is impossible.[2]

Recently, there has been a scarcity in the availability of bones for teaching in India. The demand of dry human bones for students of medicine, dentistry, physiotherapy, and allied sciences is increasing enormously. The supply is scarce. Hence, a solution must be found to meet the demand as the supply dwindles. Retrieval of the skeleton from the dissected cadavers may be a choice. Currently, underutilized sources of human skeletal material from the preserved tissue offer a solution.[2]

The retrieval of bones and removal of all soft tissue is called maceration. Various maceration techniques exist in the literature. All maceration techniques aim at removing the soft tissue from the bone. In general, several steps are required: the complete removal of all soft tissue and the processes to degrease the bone and then whiten it. Initially, as much soft tissue as possible is physically removed. This includes skinning, gutting, and usually, disarticulation in order for the bone to macerate at a more reasonable pace. In general, these techniques fall into the following categories: boiling, using chemicals or burying or using dermestid beetles.[3] Retrieval of bones from formalin-fixed cadavers is difficult. No formal standards of maceration exist. Traditional methods are tedious and time-consuming. There is a practical need for research with fixed specimens. Producing dry skeletons without bone modification from preserved tissue must be exact, as they do not macerate easily.

Therefore, a study was planned to evaluate the different methods to procure bones in terms of efficiency, effectiveness, and destructiveness. The purpose of this study is to use various methods and find the most suitable method in terms of time consumed, ease of access, and complete procurement of bones from embalmed cadavers.


  Material and Methods Top


The study was carried after taking clearance from the Institutional Ethics Committee. The study was conducted on dissected human cadavers made available from the department of anatomy, of a reputed medical college. Universal precautions were observed while handling tissue and chemicals. Four different methods were employed and were compared. For the first three methods, after dissection, the cadavers were buried in the burial section of the department for 2 years. The cadavers were excavated after 2 years. The bones were washed.

In the first method, after excavation, the bones were dipped in detergent for 24 h. Any remaining soft tissue was removed manually using a brush. The bones were washed and dried.

In the second method, bones were dipped in a quick lime solution. Then, they were washed and dried.

In the third method, after excavation, the bones were dipped in detergent for 24 h. Bones were then chemically treated with hydrogen peroxide. Two liters of 30% H2O2 were dissolved in 15 L water. The bones were dipped and left covered for 24 h. They were then washed and dried in sun.

In the fourth method, the dissected parts were defleshed. They were not buried. Bones were freed from the joint. A paste was made using 100 g baking soda and 30% H2O2. This paste was applied on the bones and left covered for 24 h. Next day, they were washed and dried. The methods were compared on the basis of time required for maceration, ease of soft-tissue removal, and complete procurement or minimal damage to bone tissue.

Observations

The following observations were made.

Method 1 use of detergent

Time to macerate – More than 2 years.

Ease of access to remove soft-tissue cleaning was effective. Bones had dark color. Soft tissue could not be removed from the ends and had to be removed manually.

Destructiveness – No damage to bone. Bones were darker.

Method 2 use of quick lime

Time to macerate – More than 2 years.

Ease of access to remove soft tissue cleaning was not effective. There was a chalky deposit on the bones [Figure 1]. The remnants of ligaments and tendons were not softened. The tissue was not removed.
Figure 1: Femur head showing chalky deposit of quick lime

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Destructiveness – No damage to bone. Soft tissue was not removed There was a deposit on the bone.

Method 3 use of hydrogen peroxide

Time to macerate – More than 2 years.

Ease of access to remove soft-tissue cleaning was effective. Bones had lighter tone. Only some of the bones needed brushing to look completely clean [Figure 2] and [Figure 3].
Figure 2: Sacrum removed after 2 years of burial

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Figure 3: Sacrum after treatment with H2O2

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Destructiveness – no damage to bone. Bones were lighter toned looked visually appealing to touch.

Method 4 use of baking soda and hydrogen peroxide – no burial

Time to macerate – The whole procedure took less than a week. In the previous three methods, the cadaver was buried the bones excavated which usually took 2 years.

Ease of access to remove soft tissue – most of the tissue comes off easily. Some tissue had to be removed manually using scalpel.

Destructiveness – no damage to bone. The bones were clean having lighter tone [Figure 4].
Figure 4: Bones treated with paste of baking soda and H2O2

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  Results Top


The advantages and disadvantages of various methods are tabulated in [Table 1].
Table 1: Showing Advantages and Disadvantages of various methods used in maceration

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Based on time required to macerate, ease of soft-tissue removal and complete procurement with minimal damage, the method using 30% H2O2 was the best. The chemicals are easily available and fairly inexpensive. For smaller specimens, the new method of applying a paste of baking soda and H2O2 was very effective. The time consumed was very less. No burial was required. Thus, we can conclude that the method using hydrogen peroxide can be used conveniently to procure full skeletons from dissected cadavers.


  Discussion Top


The need for the use of authentic osteological specimens for students as well as professionals in established fields and in new burgeoning disciplines has rapidly increased over the past several decades. Philip Tobias emphasized the importance of human skeletons for the teaching of human anatomy, including dental anatomy for students of medicine, dentistry, physiotherapy, occupational therapy, nursing, and pharmacy as well as students of comparative anatomy, primatology, and general morphology.[4] The number of new fields that rely on access to this learning material has astronomically increased and expanded. Forensics, DNA research, and molecular biology are but a few worth mentioning.[2]

To meet the increasing demand of bones in the department necessitated experimenting with various maceration procedures. In the present study, four methods were chosen for this purpose seeing the feasibility, expenses, and other factors. Although boiling of bones is time-tested procedure, it was not preferred due to obnoxious fumes and environmental hazards. Because of this disadvantage, it cannot be carried at all places.

Based on the time required to macerate, ease of soft-tissue removal, and complete procurement with minimal damage, the method using 30% H2O2 was the best. The chemicals are easily available and fairly inexpensive. Many bones can be macerated together. A single trained person can do the entire procedure. Earlier studies also concluded this method as an efficient method.[5] Burial is necessary, and bones were kept buried for 2 years. This time for burying can be reduced by burying the bones near the surface in the superficial soil.[6]

For smaller specimens, the new method of applying a paste of baking soda and H2O2 was very effective. The time consumed was very less. No burial was required. Complete knowledge and training are required to deflesh the bones and disarticulate them from the joints.

Snyder, Burdi, and Gaul introduced a method of skeletal preparation that involved a quick-acting formula they named anti-formin, prepared by combining sodium carbonate and bleaching powder.[6] Contemporary scholars caution against the guaranteed destructive properties of bleaching products.[7],[8],[9],[10] The use of bleaching powder in skeletal processing has been shown to destabilize bone structure well after the removal from the solution.[7],[9],[10] Sodium hypochlorite is effective at removing fat and whitening specimens but researchers cautioned that the resultant bone will be flaky and may crumble to dust.

As an alternative to sodium hypochlorite, hydrogen peroxide has been suggested in the literature.[8],[11] Sodium bicarbonate may be used to soften proteinaceous materials. When comparing which chemical proved the easiest to deflesh in the shortest amount of time, sodium bicarbonate came in the first place in all trials.[2],[3],[4],[5],[6],[7],[8] Thus, a combination of sodium bicarbonate and hydrogen peroxide was tried gave us fast and effective cleaning. This method requires the help of trained person to deflesh and disarticulate the bones. The reagents are easily available. It is very fast. No burial is required. This method may be difficult to be applied on the axial skeleton. It can be the method of choice for small dissected specimens.


  Conclusion Top


Maceration using 30% H2O2 gave best result New Method tried is time saving and useful for small specimens. Soft tissue was removed easily with no/minimal damage to the bone skeleton. It can be the method of choice for small dissected specimens. The methods described here are easy to perform and will help medical colleges to procure bones from preserved tissue.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Ajayi A, Edjomariegwe O, Iselaiye O. A review of bone preparation techniques for anatomical studies. Malaya J Biosci 2016;3:76-80.  Back to cited text no. 1
    
2.
Thompson MC. Preparing Skeletons for Research and Teaching from Preserved Human Specimens. MSc. East Bay: California State University; 2015.  Back to cited text no. 2
    
3.
Couse T, Connor M. A comparison of maceration techniques for use in forensic skeletal preparations. J Forensic Investig 2015;3:1-6.  Back to cited text no. 3
    
4.
Tobias PV. On the scientific, medical, dental and educational value of collections of human skeletons. Int J Anthropol 1991;6:277-80.  Back to cited text no. 4
    
5.
Modi BS, Puri N, Patnaik V. Evaluation of techniques for cleaning embalmed cadaver bones. Int J Anat Res 2014;2:810-3.  Back to cited text no. 5
    
6.
Snyder RG, Burdi A, Gaul G. A rapid technique for preparation of human fetal and adult skeletal material. J Forensic Sci 1975;20:576-80.  Back to cited text no. 6
    
7.
Fenton TW, Birkby WH, Cornelison J. A fast and safe non-bleaching method for forensic skeletal preparation. J Forensic Sci 2003;48:274-6.  Back to cited text no. 7
    
8.
Lee EJ, Luedtke JG, Allison JL, Arber CE, Merriwether DA, Steadman DW. The effects of different maceration techniques on nuclear DNA amplification using human bone. J Forensic Sci 2010;55:1032-8.  Back to cited text no. 8
    
9.
Rennick SL, Fenton TW, Foran DR. The effects of skeletal preparation techniques on DNA from human and non-human bone. J Forensic Sci 2005;50:1016-9.  Back to cited text no. 9
    
10.
Steadman DW, DiAntonio LL, Wilson JJ, Sheridan KE, Tammariello SP. The effects of chemical and heat maceration techniques on the recovery of nuclear and mitochondrial DNA from bone. J Forensic Sci 2006;51:11-7.  Back to cited text no. 10
    
11.
Aggarwal N, Gupta M, Goyal PK, Kaur J. An alternative approach to bone cleaning methods for anatomical purposes. Int J Anat Res 2016;4:2216-21.  Back to cited text no. 11
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1]



 

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  In this article
Abstract
Introduction
Material and Methods
Results
Discussion
Conclusion
References
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