欢迎访问《古脊椎动物学报》官方网站,今天是

钙化软骨中的软骨细胞比骨细胞具有更高的保存潜力?埋藏学实验初探

  • 艾莉达 ,
  • 吴倩 ,
  • 李东升 ,
  • 李志恒 ,
  • 周忠和
展开
  • 1 中国科学院古脊椎动物与古人类研究所,中国科学院脊椎动物演化与人类起源重点实验室 北京 100044
    2 中国科学院大学 北京 100049

收稿日期: 2022-12-02

  网络出版日期: 2023-03-08

基金资助

国家自然科学基金(42288201);国家自然科学基金(42172029)

Do chondrocytes within calcified cartilage have a higher preservation potential than osteocytes?A preliminary taphonomy experiment

  • M. BAILLEUL Alida ,
  • Qian WU ,
  • Dong-Sheng LI ,
  • Zhi-Heng LI ,
  • Zhong-He ZHOU
Expand
  • 1 Key Laboratory of Vertebrate Evolution and Human Origins of Chinese Academy of Sciences, Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences Beijing 100044
    2 University of Chinese Academy of Sciences Beijing 100049

Received date: 2022-12-02

  Online published: 2023-03-08

摘要

近期,在发现于美国和中国的两例早白垩世恐龙中报道了保存有细胞核物质和生物分子的软骨细胞的存在。基于多种原因,研究人员认为,钙化软骨比骨骼更有可能保存远古细胞。通过针对影响细胞保存的最主要因素:机体死亡后细胞自溶过程的停止,对这一假说进行了首次实验验证。以家鸭 (Anas platyrhynchos domesticus)为模型,比较了在不抑制自溶酶的情况下,自然埋藏60天内,软骨细胞和骨细胞的自溶过程。埋藏后的15天内,几乎所有骨细胞均发生了自溶,而钙化软骨细胞则基本未受影响。埋藏30天后,所有骨细胞均已自溶,但一些软骨细胞在埋藏60天后仍然存在。因此,即使在恶劣的条件下,钙化软骨细胞仍然能够在动物机体死亡后存活数月之久,而这一时间足以实现矿化过程。这一结果与一些法医文献中的数据相吻合,表明透明软骨细胞在机体死亡后能够长时间抵抗分解,且支持了钙化软骨比骨骼更有保存细胞结构的潜力的假说,尤其是在未能快速矿化的化石中。然而,由于所使用的标本预先经过冷冻,所观察到的自溶模式也有可能是细胞由于冰晶形成而死亡的结果,而非严格的自溶过程,因此有必要进一步对新鲜标本开展实验以提高结论的准确度。无论如何,研究明确显示透明软骨和钙化软骨的软骨细胞受到冷冻的影响比骨细胞更小。这暗示软骨细胞比骨细胞更可能在发现于永久冻土的化石或木乃伊中保存下来,而软骨(包括透明软骨和钙化软骨)可能是比骨骼更为理想的古DNA研究对象。尽管有必要对未经冷冻的新鲜样品进行后续实验并辅以DNA测序,建议古DNA专家将保存在热带和温带环境的化石中的钙化软骨作为古DNA分析的新对象。

本文引用格式

艾莉达 , 吴倩 , 李东升 , 李志恒 , 周忠和 . 钙化软骨中的软骨细胞比骨细胞具有更高的保存潜力?埋藏学实验初探[J]. 古脊椎动物学报, 2023 , 61(2) : 108 -122 . DOI: 10.19615/j.cnki.2096-9899.230309

Abstract

Chondrocytes with remnants of nuclei and biomolecules were recently reported in two Cretaceous dinosaurs from North America and China. For multiple reasons, it was hypothesized that calcified cartilage (CC) had a better potential than bone to preserve ancient cells. Here we provide the first experimental test to this hypothesis by focusing on the most important variable responsible for cellular preservation: the postmortem blockage of autolysis. We compare the timing of autolysis between chondrocytes and osteocytes in an avian model (Anas platyrhynchos domesticus) buried for up to 60 days under natural conditions that did not inhibit autolytic enzymes. Within 15 days post-burial, almost all osteocytes were already cytolyzed but chondrocytes in CC were virtually unaffected. All osteocytes were cytolyzed after 30 days, but some chondrocytes were still present 60 days post-burial. Therefore, even in harsh conditions some CC chondrocytes still survive for months postmortem on a time scale compatible with permineralization. This is consistent with other data from the forensic literature showing the extreme resistance of hyaline cartilage (HC) chondrocytes after death and does support the hypothesis that CC has a better potential than bone for cellular preservation, especially in fossils that were not permineralized rapidly. However, because the samples used were previously frozen, it is possible that the pattern of autolysis observed here is also a product of cell death due to ice crystal formation and not strictly autolysis, meaning a follow-up experiment on fresh (non-frozen samples) is necessary to be extremely accurate in our conclusions. Nevertheless, this study does show that CC chondrocytes are very resistant to freezing, suggesting that chondrocytes are likely better preserved than osteocytes in permafrost fossils and mummies that underwent a freezing-thawing cycle. It also suggests that cartilage (both hyaline and calcified) may be a better substrate for ancient DNA than bone. Moreover, even though we warrant follow-up taphonomy experiments with non-frozen samples paired with DNA sequencing, we already urge ancient DNA experts to test CC as a new substrate for ancient DNA analyses in fossils preserved in hot and temperate environments as well.

参考文献

[1] Alabbasi S F, Viramontes A C, Diaz F J et al., 2022. Loss of nuclear basophilic staining as a postmortem interval marker. Am J Foren Med Path, 43: 142-146
[2] Alberti F, Gonzalez J, Paijmans J L et al., 2018. Optimized DNA sampling of ancient bones using computed tomography scans. Mol Ecol Resour, 18: 1196-1208
[3] Alibegovi? A, 2014. Cartilage: a new parameter for the determination of the postmortem interval? J Forensic Leg Med, 27: 39-45
[4] Arkill K, Winlove C, 2008. Solute transport in the deep and calcified zones of articular cartilage. Osteoarthr Cartilage, 16: 708-714
[5] Bailleul A M, 2021. Fossilized cell nuclei are not that rare: review of the histological evidence in the Phanerozoic. Earth Sci Rev, 216: 103599
[6] Bailleul A M, Li Z H, 2021. DNA staining in fossil cells beyond the Quaternary: reassessment of the evidence and prospects for an improved understanding of DNA preservation in deep time. Earth Sci Rev, 216: 103600
[7] Bailleul A M, Zhou Z H, 2021. SEM analyses of fossilized chondrocytes in the extinct birds Yanornis and Confuciusornis: insights on taphonomy and modes of preservation in the Jehol Biota. Front Earth Sci, 9, doi: 10.3389/feart.2021.718588
[8] Bailleul A M, Zheng W, Horner J R et al., 2020. Evidence of proteins, chromosomes and chemical markers of DNA in exceptionally preserved dinosaur cartilage. Natl Sci, 7: 815-822
[9] Bell L S, Kayser M, Jones C, 2008. The mineralized osteocyte: a living fossil. Am J Phys Anthropol, 137: 449-456
[10] Briggs D, Kear A, Martill D et al., 1993. Phosphatization of soft-tissue in experiments and fossils. Geol Soc, 150: 1035-1038
[11] Brucker P U, Izzo N J, Chu C R, 2005. Tonic activation of hypoxia‐inducible factor 1α in avascular articular cartilage and implications for metabolic homeostasis. Arthritis Rheum, 52: 3181-3191
[12] Chen X, Wang W, Shang Q et al., 2009. Experimental evidence for eukaryotic fossil preservation: onion skin cells in silica solution. Precambrian Res, 170: 223-230
[13] Clark M A, Worrell M B, Pless J E, 1996. Postmortem changes in soft tissues. In: Haglund W D, Sorg M H eds. Forensic Taphonomy:the Postmortem Fate of Human Remains, 1st ed. Boca Raton, Florida: CRC Press. 1-164
[14] Cs?nge L, Bravo D, Newman-Gage H et al., 2002. Banking of osteochondral allografts, part II. Preservation of chondrocyte viability during long-term storage. Cell Tissue Bank, 3: 161-168
[15] Dettmeyer R B, 2018. Forensic Histopathology:Fundamentals and Perspectives. Giessen: Springer. 1-454
[16] Drobnic M, Mars T, Alibegovi? A et al., 2005. Viability of human chondrocytes in an ex vivo model in relation to temperature and cartilage depth. Folia Biol (Praha), 51: 103-108
[17] Frikha-Benayed D, Basta-Pljakic J, Majeska R J et al., 2016. Regional differences in oxidative metabolism and mitochondrial activity among cortical bone osteocytes. Bone, 90: 15-22
[18] Gamba C, Jones E R, Teasdale M D et al., 2014. Genome flux and stasis in a five millennium transect of European prehistory. Nat Commun, 5: 5257
[19] Genest D R, Williams M A, Greene M F, 1992. Estimating the time of death in stillborn fetuses: I. histologic evaluation of fetal organs; an autopsy study of 150 stillborns. Obstet Gynecol, 80: 575-584
[20] George J, Van Wettere A J, Michaels B B et al., 2016. Histopathologic evaluation of postmortem autolytic changes in bluegill (Lepomis macrohirus) and crappie (Pomoxis anularis) at varied time intervals and storage temperatures. PeerJ, 4: e1943
[21] Goret-Nicaise M, Dhem A, 1985. Comparison of the calcium content of different tissues present in the human mandible. Cells Tissues Organs, 124: 167-172
[22] G?therstr?m A, Collins M, Angerbj?rn A et al., 2002. Bone preservation and DNA amplification. Archaeometry, 44: 395-404
[23] Hansen H B, Damgaard P B, Margaryan A et al., 2017. Comparing ancient DNA preservation in petrous bone and tooth cementum. PLoS One, 12: e0170940
[24] Jang T H, Park S C, Yang J H et al., 2017. Cryopreservation and its clinical applications. Integr Med Res, 6: 12-18
[25] Kierdorf U, Stock S R, Gomez S et al., 2022. Distribution, structure, and mineralization of calcified cartilage remnants in hard antlers. Bone Rep, 16: 101571
[26] Lasczkowski G E, Aigner T, Gamerdinger U et al., 2002. Visualization of postmortem chondrocyte damage by vital staining and confocal laser scanning 3D microscopy. J Forensic Sci, 47: 663-666
[27] Martin D, Briggs D E, Parkes R J, 2003. Experimental mineralization of invertebrate eggs and the preservation of Neoproterozoic embryos. Geology, 31: 39-42
[28] Martin D, Briggs D E, Parkes R J, 2005. Decay and mineralization of invertebrate eggs. Palaios, 20: 562-572
[29] Murawska D, 2012. The effect of age on the growth rate of tissues and organs and the percentage content of edible and nonedible carcass components in Pekin ducks. Poult Sci, 91: 2030-2038
[30] Pallante A L, Bae W C, Chen A C et al., 2009. Chondrocyte viability is higher after prolonged storage at 37oC than at 4oC for osteochondral grafts. Am J Sports Med, 37: 24-32
[31] Paulis M, Hassan E, Abd-Elgaber N, 2016. Estimation of postmortem interval from cartilage changes of rabbit auricle. Ain-Shams J Forensic Med Clin Toxicol, 26: 61-69
[32] Pfander D, Gelse K, 2007. Hypoxia and osteoarthritis: how chondrocytes survive hypoxic environments. Curr Rheumatol Rev, 19: 457-462
[33] Pinhasi R, Fernandes D, Sirak K et al., 2015. Optimal ancient DNA yields from the inner ear part of the human petrous bone. PLoS One, 10: e0129102
[34] Pinhasi R, Fernandes D M, Sirak K et al., 2019. Isolating the human cochlea to generate bone powder for ancient DNA analysis. Nat Protoc, 14: 1194-1205
[35] Powell III J W, 2015. Multiple stain histology of skeletal fractures:healing and microtaphonomy. Ph. D thesis. Florida: University of South Florida. 1-118
[36] Prondvai E, Witten P E, Abourachid A et al., 2020. Extensive chondroid bone in juvenile duck limbs hints at accelerated growth mechanism in avian skeletogenesis. J Anat, 236: 463-473
[37] Raff E C, Villinski J T, Turner F R et al., 2006. Experimental taphonomy shows the feasibility of fossil embryos. Proc Nat Acad Sci USA, 103: 5846-5851
[38] Raff E C, Schollaert K L, Nelson D E et al., 2008. Embryo fossilization is a biological process mediated by microbial biofilms. Proc Nat Acad Sci USA, 105: 19360-19365
[39] Rogers C J, Clark K, Hodson B J et al., 2011. Postmortem degradation of porcine articular cartilage. J Forensic Leg Med, 18: 52-56
[40] Sazonova T S, Romanovsky V E, Walsh J E et al., 2004. Permafrost dynamics in the 20th and 21st centuries along the East Siberian transect. J Geophys Res Atmos, 109: 25
[41] Schweitzer M H, Zheng W, Cleland T P et al., 2013. Molecular analyses of dinosaur osteocytes support the presence of endogenous molecules. Bone, 52: 414-423
[42] Shapiro I M, Golub E E, Kakuta S et al., 1982. Initiation of endochondral calcification is related to changes in the redox state of hypertrophic chondrocytes. Science, 217: 950-952
[43] Sirak K, Fernandes D, Cheronet O et al., 2020. Human auditory ossicles as an alternative optimal source of ancient DNA. Genome Res, 30: 427-436
[44] van der Valk T, Pe?nerová P, Díez-del-Molino D et al., 2021. Million-year-old DNA sheds light on the genomic history of mammoths. Nature, 591: 265-269
[45] Williams S K, Amiel D, Ball S T et al., 2003. Prolonged storage effects on the articular cartilage of fresh human osteochondral allografts. J Bone Joint Surg, 85: 2111-2120
[46] Zheng X T, Bailleul A M, Li Z H et al., 2021. Nuclear preservation in the cartilage of the Jehol dinosaur Caudipteryx. Commun Biol, 4: 1-9
文章导航

/